Tetraselmis chuii (T. chuii) for the treatment of male infertility

Abstract

The present invention relates to a biomass of T. chuii and a pharmaceutical composition comprising the biomass of T. chuii for use in treating infertility in a male subject. The present invention also relates to a method for improving semen quality in a male subject who has normal semen, does not have oligospermia, and does not have a disorder characterized by a high percentage of sperm DNA fragmentation (SDF), comprising administering a biomass of T. chuii.

Description

Technical Field

[0001] The present invention relates to the field of compositions for use in the treatment of male infertility associated with deficiencies in semen, wherein said compositions are obtained from microalgae.

Background Art

[0002] Infertility has been diagnosed in 12 - 15% of sexually active couples. In 50% of said couples, infertility is caused by a disorder in the male partner, either alone or in combination with a disorder in the female partner.

[0003] Defects causing male infertility can be classified as incomplete sperm delivery due to disorders related to the hypothalamus or pituitary gland, the testes, or the penis or related genitalia. Deficiencies in semen parameters are often observed in infertile men. Thus, male infertility is often diagnosed by analysis of a semen sample from a male subject.

[0004] Semen parameters that generally vary in infertile men include the volume of the semen sample, the approximate number of total sperm cells, sperm motility or progressive motility, the percentage of sperm with DNA fragmentation (SDF), the number of dead sperm cells in the semen, or the presence of white blood cells in the semen. Changes in said parameters are classified within different disorders, such as oligospermia (very low sperm count), azoospermia (absence of semen), hypospermia (low semen volume), aspermia (absence of sperm in the semen), teratospermia (high morphological defects in sperm), asthenozoospermia (low sperm motility), necrospermia (high number of dead sperm), pyospermia (presence of a high amount of white blood cells in the semen), and high SDF percentage (high percentage of sperm showing DNA fragmentation).

[0005] Treatment of male infertility generally aims to treat specific physiological changes that are probably the most likely causes of the disorder. For example, surgery can be used when male infertility is caused by a blockage in the sperm transport system, such as after vasectomy. Vasectomy can generally be surgically reversed in up to 85 percent of cases, however, in many cases, men remain infertile even after the success of the intervention. When male infertility is due to low testosterone production, hormonal treatment can be administered to restore fertility (luteinizing hormone LH and follicle-stimulating hormone FSH). However, long-term treatment is necessary for sperm production to reach levels associated with male fertility.

[0006] Couples in which the male partner has been diagnosed with male infertility are often advised to follow different reproductive assistance methods. Such invasive methods include artificial insemination (IUI), in vitro fertilization (IVF), or IVF with intracytoplasmic sperm injection (ICSI).

[0007] Therefore, independent of the cause of the disorder, there is a need in the art for non-invasive and effective treatments for male infertility. SUMMARY OF THE INVENTION

[0008] The inventors have observed that administration of microalgae from the species Tetraselmis chuii (T. chuii) to male subjects having a disorder associated with male infertility and characterized by changes in specific semen parameters, such as idiopathic oligospermia, asthenozoospermia, and / or teratozoospermia, results in normalization of the corresponding semen parameters and thus contributes to the treatment of the disorder.

[0009] In particular, as shown in Examples 1-4 and Figures 1-8, the inventors observed that in semen samples of subjects with idiopathic oligospermia, asthenozoospermia, and / or teratozoospermia, the levels of semen volume, sperm concentration, total sperm count per ejaculate, and parameters of progressive motility increased from levels below the corresponding lower reference level (LRL) to levels above the LRL upon administration of T. chuii. Additionally, as shown in Example 5 and Figure 10, the inventors observed that after administration of T. chuii, the percentage of subjects analyzed to exhibit teratozoospermia decreased from 35% to 10%, indicating that the percentage of morphologically normal sperm in the semen samples of said patients increased to a value above the corresponding LRL. Finally, as shown in Example 6 and Figure 11, the inventors observed that after administration of T. chuii, 30% of the patients originally diagnosed with idiopathic oligospermia, asthenozoospermia, and / or teratozoospermia had normal semen.

[0010] Accordingly, in a first aspect, the present invention relates to the biomass of T. chuii or a pharmaceutical composition comprising the biomass of T. chuii for use in the treatment of infertility in male patients.

[0011] In a second aspect, the present invention relates to a method for improving the quality of semen of a male subject having normal semen, not having oligospermia, and not having a disorder characterized by a percentage of sperm DNA fragmentation (SDF) above a reference value, the method comprising the step of treating the subject by administering the biomass of T. chuii.

[0012] In a third aspect, the present invention relates to the use of the biomass of T. chuii as an oral supplement for improving the quality of semen of a male subject having normal semen, not having oligospermia, and not having a disorder characterized by a percentage of SDF above a reference value. BRIEF DESCRIPTION OF THE DRAWINGS

[0013]

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Mode for Carrying Out the Invention

[0014] I - Medical Use of the Present Invention In a first aspect, the present invention relates to a biomass of Tetraselmis chuii (T. chuii), a protein extract of T. chuii, or a pharmaceutical composition comprising a biomass of T. chuii or a protein extract of T. chuii for use in the treatment of infertility in male patients.

[0015] 「 Tetraselmis chui( Tetraselmis chuii ) The term "", more generally "Tetraselmis chuii" or " T. chuii " as written, when used herein, Chlorodendron algae ( Chlorodendrophyceae ) class, Chlorodendron ( Chlorodendrales ) order, Chlorodendron ( Chlorodendraceae ) refers to marine unicellular algae (microalgae) belonging to the family, which are green, motile, and usually grow to a length of 10 μm × width of 14 μm. In certain embodiments, T. chuii corresponds to the organism identified by Taxonomy ID: 63592 (NCBI: txid63592) in the NCBI database.

[0016] " biomass " as used herein refers to biological materials including living organisms or recently living organisms. Thus, this term includes not only biological materials or organic substances that make up an organism, but also biological materials or organic substances that occur in spontaneous or non-spontaneous (i.e., induced) biological processes. As will be understood by those skilled in the art, Biomass of T. chuii is the biomass as precisely defined, where the organism is the alga T. chuii. Generally, the said biomass is obtained from a sample of T. chuii.

[0017] In certain embodiments, the biomass of T. chuii is obtained from one cell of T. chuii. In another specific embodiment, the biomass of T. chuii is obtained from several cells of T. chuii. Thus, in a preferred embodiment, the biomass of T. chuii is obtained from a cell culture of T. chuii.

[0018] In certain embodiments, the cell culture of T. chuii from which the biomass of T. chuii is obtained is performed under conditions suitable for the growth of the microalgae T. chuii, which are well known to those skilled in the art. As is well known to those skilled in the art, said conditions are characterized, for example, by the use of a suitable medium such as the F / 2 culture medium [Guilard R.R.L. and Ryther, J.H. 1962. "Studies of marine planktonic diatoms. I. Cyclotela nana Hustedt and Detonula confervaceae (Cleve) Gran." Can. J. Microbiol. 8, 229-239], performance under controlled conditions of sunlight or suitable lighting conditions (light intensity), as well as pH, temperature, and carbon dioxide (CO2) supply. The F / 2 culture medium contains, in an aqueous medium, a nitrogen source, a phosphorus source, trace metals such as, for example, sodium, iron, copper, zinc, cobalt, manganese and molybdenum, and a mix of vitamins such as, for example, cyanocobalamin (vitamin B12), thiamine (vitamin B1) and biotin.

[0019] The light intensity is adjusted so that photosynthesis is possible, and thus, this can vary within a wide range in certain embodiments, but the light intensity applied to the culture medium is between 1 and 2000 μmol photons m -2 s -1 of photosynthetically active radiation (PAR) outdoors, and typically about 150 μmol photons m -2 s -1 indoors. The pH can usually vary between about 7 and about 8.5 and can typically be about 7.5.

[0020] The temperature that promotes the growth of T. chuii is usually selected to be between about 17°C and about 28°C, typically including between about 24°C and 26°C.

[0021] The culture is typically carried out with aeration, for example, with approximately 0.5 - 5%, preferably about 1 - 2% CO2 in ambient air, with or without aeration.

[0022] In a preferred embodiment, the cultivation of T. chuii is carried out under standard conditions, using an F / 2 culture medium, at 150 μmol photons m -2 s -1 (PAR), at a temperature between 24 °C and 26 °C, preferably at 25 °C, at pH 7.5, using 1-2% CO2-enriched atmospheric air.

[0023] In a more preferred embodiment, the cell culture of T. chuii from the biomass of T. chuii is carried out in an outdoor tubular photobioreactor using an F / 2 culture medium, under natural light cycles, at ambient temperature, and at a pH controlled by CO2 injection. Specifically, the cell culture is carried out using an F / 2 culture medium, at a light intensity between 1 and 2000 μmol photons m -2 s -1 (PAR), at a temperature between 10 °C and 35 °C, at a pH between 7 and 8.5, and at a salinity of 35 practical salinity units (PSU). In addition, this was carried out using a 12:12 light cycle (i.e., 12 hours of light and 12 hours of darkness).

[0024] In another specific embodiment, the cell culture of T. chuii from which the biomass of T. chuii is obtained is carried out under abiotic stress conditions.

[0025] As used herein, the expression " abiotic stress " relates to the negative impact of abiotic factors on organisms surviving in a particular environment. The abiotic variables must affect the environment in a significant way beyond their normal range of variation that is harmful to the performance of a population of organisms or individual physiological functions.

[0026] There are several abiotic stress factors that can affect the growth of microalgae, such as T. chuii, as well as the production of compounds and their metabolites. However, in a particular embodiment, the abiotic stress applied to the cell culture from which the biomass of T. chuii for the medical use of the present invention is obtained is selected from the group consisting of high redox potential, high temperature, high salinity, and nitrogen starvation.

[0027] According to the present invention, abiotic stress based on a high redox potential includes maintaining a culture medium having a redox potential of at least 100 mV, at least 200 mV, at least 300 mV, at least 400 mV, at least 500 mV, at least 600 mV, at least 700 mV, at least 800 mV, at least 900 mV, at least 1000 mV, and the abiotic stress can be obtained by conventional methods for obtaining a high redox potential state in a culture medium, such as by adding ozone that normally occurs using an ozone generator by the reaction of air and UV. The amount of ozone added is required to achieve and maintain a culture medium having a redox potential of at least 100 mV, at least 200 mV, at least 300 mV, at least 400 mV, at least 500 mV, at least 600 mV, at least 700 mV, at least 800 mV, at least 900 mV, at least 1000 mV.

[0028] According to the present invention, abiotic stress based on a high temperature includes maintaining a culture medium at a temperature of at least 28°C, at least 30°C, at least 35°C, at least 40°C, at least 45°C, at least 50°C, and the temperature in the culture medium can be obtained by conventional methods. Since the culture can be indoors or outdoors, for indoor culture, the temperature of the room to be cultured must be set above 28°C, while for outdoor culture, the culture should be carried out at the appropriate location where the temperature naturally reaches the above temperature, for example, by the environmental temperature in southern Spain such as Cadiz, Malaga, Seville, etc., for example, during spring and summer.

[0029] According to the present invention, abiotic stress based on high salinity conditions includes maintaining the culture medium at a salinity of at least 35 PSU (practical salinity unit), at least 40 PSU, at least 45 PSU, at least 50 PSU, at least 100, at least 200, at least 300. Those skilled in the art know how to determine the salinity of the culture medium by using standard techniques (UNESCO, 1981, "Background papers and supporting data on the practical salinity scale 1978", Unesco Technical papers in marine science, 37). Abiotic stress based on high salinity conditions can be easily obtained by adding salt to the culture medium until the salinity conditions are reached, for example, by evaporating natural seawater until the target salinity is reached, or by adding commercially available sea salt.

[0030] According to the present invention, abiotic stress based on nitrogen starvation includes growing T. chuii under conditions of nitrogen deficiency, limitation or exhaustion, and said abiotic stress can be easily achieved by stopping the supply of nitrogen to the culture medium (i.e., by discontinuing the addition of nitrogen to the culture medium).

[0031] In a preferred embodiment, the abiotic stress includes nitrogen starvation. The term " nitrogen starvation ", as used herein, refers to conditions such that the supply of nitrogen is such that the nitrogen concentration in the culture medium is less than 200 μm, less than 100 μm, less than 10 μm, less than 0.1 μm, less than 0.001 μm.

[0032] Methods for obtaining the biomass of T. chuii are well known to those skilled in the art. Non-limiting examples of said methods include filtration or centrifugation of a cell culture or a group of T. chuii cells from which biomass is obtained. The collected biomass is then preferably and advantageously washed to remove non-biological materials (such as mineral salt precipitates, etc.).

[0033] In a preferred embodiment, the biomass referred to in the first aspect of the present invention, or the biomass contained in the pharmaceutical composition referred to in the above aspect of the present invention, is fresh or dehydrated. In another preferred embodiment, the biomass is dehydrated. In one embodiment, the biomass is fresh.

[0034] 「 dehydrated biomass 」 or 「 lyophilized biomassThe expression "」, when used in this specification, refers to the biomass as defined in this specification, where water has been completely or partially removed. In a preferred embodiment, the water has been completely removed. In another specific embodiment, the water content removed corresponds to at least 40%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 88.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, preferably at least 70%, more preferably at least 75% of the weight of the fresh biomass. Methods for determining the percentage of water removed in the dehydrated biomass are well known to those skilled in the art and include determining the weight of a given biomass before and after dehydrating the biomass, where the difference is the weight of the amount of water removed during dehydration. Methods for dehydrating the biomass are also well known to those skilled in the art and include sun drying, convective drying, spray drying, freeze drying, roller / drum drying, fluidized bed drying, vacuum tray drying, and / or solar conductive drying (Chen et al., 2015. Dewatering and drying methods for microalgae. Drying Technology 33(4):443-454.; Bheda et al., 2018. Drying of algae by various drying methods. IDS’2018 - The 21st International Drying Symposium, pp. 1791 - 1797. Valencia, Spain, September 11 - 14, 2018, Editorial Universitat Politecnica de Valencia).

[0035] 「 fresh biomassAs used herein, the expression " refers to non-dehydrated biomass, i.e., biomass in which the water contained in the biomass has not been partially or completely removed. In a preferred embodiment, fresh biomass is directly obtained by any of the methods provided above for obtaining biomass.

[0036] 「 Protein extract of T. chuii As used herein, the expression " refers to a sample of protein obtained upon extraction of protein from T. thuyii cells. More specifically, the protein extract is obtained from a sample of T. thuyii. In a preferred embodiment, the sample of T. thuyii from which the protein extract is obtained is a sample of T. thuyii from which the biomass of T. thuyii is obtained and which is defined in any of the above embodiments. Thus, a cell culture from a sample of T. thuyii from which a protein extract of T. thuyii is obtained is also a sample from which a sample for obtaining the biomass of T. thuyii is obtained. Therefore, a cell culture of T. thuyii from a sample of T. thuyii from which a protein extract of T. thuyii is obtained is a cell culture defined in any of the above embodiments, i.e., the conditions of said cell culture are any of the conditions defined above.

[0037] The methods used to obtain a protein extract of T. chuii from a sample of T. chuii are well known to those skilled in the art. Non-limiting examples of such methods include the addition of an extraction buffer to the T. chuii biomass obtained as described above, followed by cell lysis, centrifugation of the sample, and isolation of the supernatant of the centrifuged sample. Protein extraction buffers are well known to those skilled in the art. Non-limiting examples of protein extraction buffers include phosphate buffers, such as 220 mM KH2PO4 buffer at pH 7.8. Methods for lysing cells are well known to those skilled in the art and include the use of ultrasound at several intervals, such as 4 cycles of ultrasound at 20% amplitude for 30 seconds each with a 10-second break between each cycle. Other non-limiting examples of methods for obtaining a protein extract from a sample of T. chuii are described in Bleakley S. and Hayes M. 2017. Foods 6(5):33.

[0038] 「 pharmaceutical composition 」, as used herein, refers to a composition comprising a therapeutically effective amount of the biomass of T. chuii or a protein extract of T. chuii, and at least one pharmaceutically acceptable excipient.

[0039] The pharmaceutical composition according to the present invention can be prepared, for example, as an injection such as a liquid solution, suspension, and emulsion. 「 pharmaceutically acceptable excipient 」, or 「 pharmaceutically acceptable carrier 」, 「 pharmaceutically acceptable diluent 」, or 「 pharmaceutically acceptable vehicleThe term "」 is used interchangeably herein and refers to a non-toxic, solid, semi-solid or liquid filler, diluent, encapsulating material or any conventional type of pharmaceutical adjuvant. A pharmaceutically acceptable carrier is essentially non-toxic to the recipient at the dosages and concentrations employed and is compatible with the other ingredients of the formulation. Suitable carriers include, but are not limited to, water, dextrose, glycerol, normal saline, ethanol, and combinations thereof. The carrier can contain additives such as wetting or emulsifying agents, pH buffering agents, or adjuvants that enhance the effectiveness of the formulation. Adjuvants can be selected from the group consisting of sterile liquids such as water, and oils of animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water or normal saline aqueous solutions and aqueous dextrose and glycerol solutions are preferably used as vehicles, especially for injectable solutions. Suitable pharmaceutical vehicles are described in "Remington’s Pharmaceutical Sciences", 21st Edition, 2005, by E.W. Martin.

[0040] 「 therapeutically effective amount The term "」 when used herein, in relation to the biomass or protein extract of T. Chuii, for providing a desired effect, i.e., derived from a disease one or more symptomRegarding the sufficient amount of the biomass or protein extract to achieve appropriate prevention, cure, delay, reduction of its severity, or alleviation, generally, among other causes, it is determined by the characteristics of the agent itself (i.e., the biomass or protein extract of T. tuyi) and the achieved therapeutic effect. This also depends on the subject being treated, the severity of the disease the subject is suffering from, the dosage form selected, etc. For this reason, the dosages that can be cited in the present invention should only be considered as a guide for those skilled in the art who must adjust the dosage according to the variable factors described above. In embodiments, the effective amount results in the alleviation of one or more symptoms of the disease being treated. Even though individual needs vary, the determination of the optimal range for the therapeutically effective amount of the compounds according to the present invention belongs to the general experience of those skilled in the art. Generally, the dosage required to provide an effective treatment that can be adjusted by those skilled in the art varies according to age, health, fitness, gender, diet, weight, degree of receptor variation, frequency of treatment, nature and condition of the injury, nature and degree of the dysfunction or disease, medical condition of the subject, route of administration, activity, effectiveness, pharmacokinetics, and toxicity profile of the specific compound used, whether systemic drug delivery is used, and whether the compound is administered as part of a drug combination.

[0041] In certain embodiments, the pharmaceutical composition for the medical use of the present invention comprises the aforementioned biomass of T. tuyi. In another certain embodiment, the pharmaceutical composition for the medical use of the present invention comprises the aforementioned protein extract of T. tuyi.

[0042] In a preferred embodiment, the male patient of the first aspect of the present invention is characterized in that the subject exhibits male infertility.

[0043] “ infertility ” As used herein, refers to a genital disorder defined by failure to achieve a clinical pregnancy after 12 months or more of regular unprotected sexual intercourse. “ male infertilityThe expression " ", when used in this specification, refers to infertility when diagnosed in a male subject. Male infertility can result from a very diverse range of causes. Common causes of male infertility are changes in one or several semen parameters, such as the volume of the semen sample, the approximate number of total sperm cells, sperm motility or progressive motility, the percentage of sperm with DNA fragmentation, the number of dead sperm cells in the semen, or the concentration of white blood cells in the semen. Changes or deficiencies in one or several semen parameters can lead to different disorders, which can be classified as oligospermia, azoospermia, hypospermia, aspermia, teratospermia, asthenospermia, necrospermia, pyospermia, high SDF in the semen or high oxidative stress (which can lead to an increase in the percentage of sperm with 8-hydroxy,2'-deoxyguanosine, which is associated with high static redox potential levels). Additionally, disorders characterized by a combination of two or more of the above disorders, such as in the case of asthenoteratospermia, astheno-oligoteratospermia, oligo-teratospermia, or astheno-oligospermia, can also occur. The changes in semen parameters observed in these disorders can be due to known causes, however, in a high percentage of cases, the cause of the changes is unknown and is therefore considered an idiopathic disorder.

[0044] " idiopathic The expression " " when used in this specification refers to a disease or disorder understood as any disease or disorder due to unknown causes or of apparent spontaneous origin. Thus, the term "idiopathic" when applied to a disease or disorder described in this specification, such as male infertility, oligospermia, asthenospermia, teratospermia, or high SDF, refers to the corresponding disease or disorder when the cause from which it derives is unknown.

[0045] In certain embodiments, the infertility is idiopathic infertility.

[0046] " semen " or " seminal plasmaAs used herein, the term " ejaculation " refers to the body fluid produced and released by the male genitalia in a healthy subject, which contains sperm cells. In humans, semen contains, in addition to sperm cells, various enzymes and fructose. These elements not only promote the survival of sperm cells but also provide a medium for them to swim. The process by which semen is released is called " ejaculate ", and the released semen is known as "

[0047] As sperm cells pass through the ejaculatory ducts, they are mixed with body fluids mainly derived from the seminal vesicles, prostate gland, and bulbourethral glands. The typical components of semen are as follows: sperm produced and released from the testes (2 - 5%), seminal vesicle secretions (65 - 75%, i.e., composed of amino acids, citrate, flavin, fructose, and certain enzymes), prostate gland secretions (25 - 30%, i.e., composed of acid phosphatase, citrate, fibrinolysin, proteolytic enzymes, zinc, etc.), and bulbourethral gland secretions (<1%, i.e., containing galactose and mucus). sperm The term "

[0048] " refers to multiple sperm cells or sperm contained in semen. sperm cell " or the term " sperm ", as used herein, refers to a mature male germ cell that can fertilize a mature egg (secondary oocyte) in sexual reproduction. Sperm develop in the seminiferous tubules of the testes. It is formed by a head and a tail, where the tail is composed of a midpiece, a principal piece, and an end piece. The head contains a nucleus with tightly coiled chromatin fibers surrounded by a flat sac called the acrosome, which contains enzymes involved in the penetration of the oocyte. The tail or flagellum provides motility to the sperm. The neck or midpiece contains mitochondria that provide energy for the motility of the sperm during ATP production. The characteristics of the morphologically normal head, midpiece, and principal piece of sperm are provided in the definition of "morphologically normal sperm" below. The developmental stages of sperm are spermatogonia, spermatocytes, spermatids, and finally sperm.

[0049] The expression " semen sample " when used in this specification refers to a sample containing or consisting of semen from a male patient. In a preferred embodiment, the sample comprises semen from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 ejaculations from a male patient, preferably 2 ejaculations from a male patient, more preferably 1 ejaculation from a male patient. In an even more preferred embodiment, the semen sample consists of semen from any one of several ejaculations immediately upon instruction, preferably semen from 1 ejaculation from a male patient. Thus, in a preferred embodiment, the volume of semen in a sample of a patient's or subject's semen is equal to the volume of the ejaculate of said patient or subject. In another preferred embodiment, the volume of a sample of a patient's or subject's semen is the volume of the ejaculate of said patient or subject. In addition, the semen parameters in a sample of a patient's or subject's semen correspond to the semen parameters in the ejaculate of the semen of said patient or subject. The ejaculate is what is contained in the semen sample. Examples of the semen parameters include total sperm count, sperm concentration, semen volume, percentage of morphologically normal sperm, percentage of progressively motile sperm, percentage of viable sperm, and percentage of immotile sperm. In one embodiment, a sample of a patient's or subject's semen refers to the semen of the ejaculate of said patient or subject, and vice versa. Reference values for semen parameters are described herein with respect to the semen of the ejaculate. Thus, in certain embodiments, the reference values for semen parameters correspond to the reference values for semen parameters in a semen sample. Methods for obtaining and storing semen samples are well known to those skilled in the art and include methods for collecting semen samples as described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition.

[0050] In a preferred embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is obtained at least 1 day after abstinence, at least 2 days after abstinence, at least 3 days after abstinence, at least 4 days after abstinence, at least 5 days after abstinence, at least 6 days after abstinence, at least 7 days after abstinence, at least 8 days after abstinence, at least 9 days after abstinence, at least 10 days after abstinence, at least 15 days after abstinence, at least 20 days after abstinence, at least 25 days after abstinence, at least 30 days after abstinence. In a more preferred embodiment, any of the samples is obtained at least 2 days after abstinence.

[0051] In another preferred embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is obtained less than 5 days after abstinence, less than 6 days after abstinence, less than 7 days after abstinence, less than 8 days after abstinence, less than 9 days after abstinence, less than 10 days after abstinence, less than 11 days after abstinence, less than 15 days after abstinence, less than 20 days after abstinence, less than 25 days after abstinence, less than 30 days after abstinence. In a more preferred embodiment, any of the samples is obtained less than 7 days after abstinence.

[0052] In a particular embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is at least 2 days after abstinence, but less than 5 days after abstinence, less than 6 days after abstinence, less than 7 days after abstinence, less than 8 days after abstinence, less than 9 days after abstinence, less than 10 days after abstinence, less than 11 days after abstinence, less than 15 days after abstinence, less than 20 days after abstinence, less than 25 days after abstinence, less than 30 days after abstinence, preferably less than 7 days after abstinence.

[0053] In another particular embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is at least 3 days after abstinence, but less than 5 days after abstinence, less than 6 days after abstinence, less than 7 days after abstinence, less than 8 days after abstinence, less than 9 days after abstinence, less than 10 days after abstinence, less than 11 days after abstinence, less than 15 days after abstinence, less than 20 days after abstinence, less than 25 days after abstinence, less than 30 days after abstinence, preferably less than 7 days after abstinence.

[0054] In another specific embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is after at least 4 days of abstinence, but less than 5 days later, less than 6 days later, less than 7 days later, less than 8 days later, less than 9 days later, less than 10 days later, less than 11 days later, less than 15 days later, less than 20 days later, less than 25 days later, less than 30 days later, and preferably is obtained less than 7 days after abstinence.

[0055] In another specific embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is after at least 5 days of abstinence, but less than 6 days later, less than 7 days later, less than 8 days later, less than 9 days later, less than 10 days later, less than 11 days later, less than 15 days later, less than 20 days later, less than 25 days later, less than 30 days later, and preferably is obtained less than 7 days after abstinence.

[0056] In another specific embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is after at least 6 days of abstinence, but less than 7 days later, less than 8 days later, less than 9 days later, less than 10 days later, less than 11 days later, less than 15 days later, less than 20 days later, less than 25 days later, less than 30 days later, and preferably is obtained less than 7 days after abstinence.

[0057] In another specific embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is after at least 7 days of abstinence, but less than 8 days later, less than 9 days later, less than 10 days later, less than 11 days later, less than 15 days later, less than 20 days later, less than 25 days later, less than 30 days later, and preferably is obtained less than 7 days after abstinence.

[0058] In a preferred embodiment, any of the semen samples referred to in any of the embodiments of any aspect of the present invention is after at least 2 days of abstinence, but less than 7 days later.

[0059] 「 abstinenceAs used herein, the expression "no ejaculation by a male subject during a period" refers to the absence of ejaculation by a male subject during a period. In a preferred embodiment, any semen sample of any aspect of the invention is obtained from one or more ejaculates from a male subject, preferably from a single ejaculate from a male subject. Thus, in another preferred embodiment, abstinence, as used herein, also refers to the absence of obtaining (i.e., collecting) a sample of semen from the male patient.

[0060] 「 treatment As used in a first aspect of the invention, the term "treatment" refers to any type of treatment that is aimed at terminating, preventing, alleviating, or reducing susceptibility to a clinical condition described herein. In a preferred embodiment, the term "treatment" relates to a prophylactic treatment of a disorder or condition as defined herein (i.e., a treatment to reduce susceptibility to a clinical condition). Thus, "treatment", "treating" and their equivalent terms refer to obtaining a desired pharmacological or physiological effect, and include any treatment of a pathological condition or disorder in mammals, including humans. The effect may be prophylactic in terms of completely or partially preventing the disorder or its symptoms, and / or therapeutic in terms of partially or completely curing the disorder and / or the adverse effects caused by the disorder. That is, "treatment" includes (1) preventing a disorder from occurring or recurring in a subject, (2) inhibiting the disorder, e.g., arresting its development, (3) stopping or terminating the disorder, or at least the symptoms associated therewith, such that the host no longer suffers from the disorder or its symptoms, e.g., restoring or repairing a lost, damaged, or defective function, or stimulating an inefficient process, e.g., causing regression of the disorder or its symptoms, or (4) alleviating, reducing, or ameliorating the disorder, or the symptoms associated therewith, where ameliorating is used in a broad sense to refer to at least a reduction in the magnitude of the change in the parameters associated with the disorder.

[0061] In a particular embodiment, the treatment referred to in the first aspect of the invention comprises the administration of a T. chuui biomass, a T. chuui protein extract, or a pharmaceutical composition as defined herein. The T. chuui biomass, T. chuui protein extract, or pharmaceutical composition for use according to the invention can be administered to a subject by any suitable route of administration, for example parenterally (e.g., intramuscularly, intravenously, subcutaneously, nasally, etc.), enterally (i.e., orally, rectally, etc.), topically, etc. In a preferred embodiment, the T. chuui biomass, T. chuui protein extract, or pharmaceutical composition referred to in the first aspect of the invention is administered orally.

[0062] " subject ", patient "or" individual The term "subject" is used interchangeably herein to refer to any member of the animal kingdom, and may be a vertebrate, such as a fish, bird, reptile, amphibian, or mammal, such as a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, bird, cat, guinea pig, or rodent. Preferably, the subject is a mammal, more preferably a human.

[0063] In certain embodiments, the subject has oligospermia. In more particular embodiments, the subject has idiopathic oligospermia.

[0064] " oligospermia The term "as used herein" refers to a disorder in a male subject or patient characterized in that the semen of a male patient exhibits a total number of sperm per ejaculate, or a sperm concentration, below the LRL established by the WHO (Human Sperm Examination and Procedures WHO Laboratory Manual, 2010, 5th ed.).

[0065] The term "total sperm count" or "total sperm count" as used herein refers to the total number of sperm per ejaculate.

[0066] The term "sperm concentration" or "spermatozoa concentration", as used herein, refers to the total number of sperm per ejaculate volume divided by the volume of the ejaculate.

[0067] In a preferred embodiment, if a male subject has oligospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the male subject's semen must show a total sperm count and / or sperm concentration per ejaculate that is below the reference value.

[0068] The term "reference value", as used herein, relates to a predetermined criterion used as a reference for evaluating a value or data obtained from a sample collected from a subject. The reference value or reference level can be an absolute value, a relative value, a value having an upper or lower limit, a range of values, an average value, a median value, a mean value, or a value compared to a specific control or baseline value. The reference value can be based, for example, on the value of an individual sample such as a value obtained from a sample from the subject being tested at an earlier time point. The reference value can be based, for example, on a number of samples from a control population of age-matched groups over time, or on a pool of samples including or excluding the sample being tested. In a particular embodiment, the reference value is obtained from one or more male subjects, preferably healthy subjects, more preferably male subjects considered to be fertile.

[0069] In a particular embodiment, the reference value for oligospermia is the LRL established by the WHO ("WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition). The WHO states in the manual that the LRL for total sperm count is 39×10 6 per ejaculate and the LRL for sperm concentration is 15×10 6It is established as an individual. Methods for determining the total number of sperm per ejaculate, or the concentration of sperm in the semen of a male subject or a semen sample of a male subject, are well known to those skilled in the art. Non-limiting examples of such methods include those described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for counting sperm in a semen sample, or for determining the number of sperm in a semen sample, particularly the methods described in Sections 2.7 - 2.8 of said document.

[0070] In certain embodiments, the subject has azoospermia. In more specific embodiments, the subject has idiopathic azoospermia.

[0071] 「 aspermia 」The term, as used herein, refers to a disorder characterized by a male subject not ejaculating semen. This disorder can be due to the absence of ejaculate or retrograde ejaculation. Methods for determining whether a male subject exhibits azoospermia are well known to those skilled in the art and include methods that are part of the common general knowledge for determining the inability of a subject to ejaculate semen under any conditions. Methods for determining retrograde ejaculation are described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2020, 5th edition.

[0072] In certain embodiments, the subject has hypospermia. In more specific embodiments, the subject has idiopathic hypospermia.

[0073] 「 hypoospermia 」The term, as used herein, refers to a disorder characterized by an abnormally low volume of semen per ejaculate in a male subject, i.e., below the LRL.

[0074] The term "volume of semen", as used herein, refers to the volume of semen per ejaculate.

[0075] In a preferred embodiment, when a male subject has oligospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 ejaculates of the male subject must show a semen volume below the reference value. In certain embodiments, the reference value is the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition. In certain embodiments, when a male subject has oligospermia and the semen sample of the male subject contains semen from only 1 ejaculate, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples from the male subject must show a semen volume per ejaculate below the reference value, where the reference value is preferably the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition. The LRL for the semen volume of ejaculates established by the WHO in the Manual is 1.5 ml. Methods for determining the semen volume of ejaculates, or the semen volume per ejaculate of a semen sample, are well known to those skilled in the art. Non-limiting examples of such methods include the methods for sample collection and for measuring the volume of semen samples described in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, particularly Sections 2.2 and 2.3.4 of said document.

[0076] In certain embodiments, the subject has azoospermia. In more particular embodiments, the subject has idiopathic azoospermia.

[0077] 「 azoospermiaAs used herein, the term " " refers to a disorder characterized by the semen of a male subject being given as the number of sperm below the limit of quantification for the evaluation method used, and / or the absence of sperm in the testicular semen and / or in the ejaculated semen. In a preferred embodiment, when a male patient has azoospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject must be shown to have no sperm given as the number of sperm below the limit of quantification for the evaluation method used. Methods for determining the absence of sperm in the semen of a male patient or in a sample of the semen of a male patient are well known to those skilled in the art. Non-limiting examples of such methods include those described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for determining the number of sperm in a semen sample and for determining the number of sperm in a semen sample with a low sperm count, in particular, the methods described in Sections 2.7 to 2.11.

[0078] In certain embodiments, the subject has teratozoospermia. In more particular embodiments, the subject has idiopathic teratozoospermia.

[0079] 「 teratozoospermia As used herein, the term " " refers to a disorder characterized by the semen of a male subject showing the percentage of morphologically normal sperm below the LRL.

[0080] As used herein, the term "percentage of morphologically normal sperm" refers to the percentage of morphologically normal sperm relative to the total number of sperm per ejaculate.

[0081] In a preferred embodiment, when a male patient has teratozoospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject show a percentage of morphologically normal sperm below the reference value. In certain embodiments, the reference value is the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition. The WHO has established in said manual the LRL for the percentage of morphologically normal sperm as 4%. Methods for determining the percentage of morphologically normal sperm in the semen of a male subject or in a semen sample of a male subject are well known to those skilled in the art. Non-limiting examples of said methods include the methods described in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for determining the percentage of morphologically normal sperm in a semen sample, in particular the methods described in sections 2.13 to 2.17 of said document.

[0082] 「 morphologically normal sperm When used in this specification, the expression "refers to the following sperm. i) The head is smooth, has a regular contour, and is generally oval-shaped. This includes an acrosome region with a distinct contour that includes 40 - 70% of the head region. The acrosome region does not contain large vacuoles, does not contain more than two small vacuoles, and this does not occupy more than 20% of the sperm head. The post-acrosomal region does not contain any vacuoles. ii) The mid-piece is slender, regular, and approximately the same length as the sperm head. The main axis of the mid-piece is aligned with the main axis of the sperm head. Residual cytoplasm is considered abnormal only if it is excessive, i.e., more than one-third of the sperm head size. iii) The principal piece has a uniform diameter along its length, which is thinner than the mid-piece and is approximately 45 μm in length (about 10 times the length of the head). This may loop back on itself provided there are no acute angles indicating flagellar breakage.

[0083] In certain embodiments, the subject has asthenozoospermia. In more particular embodiments, the subject has idiopathic asthenozoospermia.

[0084] 「 asthenozoospermia 」 As used herein, the term refers to a disorder characterized in that the semen of a male subject exhibits a percentage of progressively motile (PR) sperm below the LRL.

[0085] As used herein, the term "percentage of progressively motile sperm" refers to the percentage of progressively motile sperm relative to the total number of sperm per ejaculate.

[0086] In preferred embodiments, when a male patient has asthenozoospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject exhibit a percentage of progressively motile (PR) sperm below the reference value. In certain embodiments, the reference value is the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition. The WHO has established the LRL for PR as 32% in said manual. Methods for determining the percentage of PR sperm in the semen of a male subject or in a semen sample of a male subject are well known to those skilled in the art. Non-limiting examples of said methods include the methods described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for determining the percentage of sperm with PR in a semen sample, in particular the methods described in section 2.5 of said document. 「 progressive motility sperm 」 As used herein, the expression refers to sperm that move actively, either linearly or in large circles, regardless of speed.

[0087] In certain embodiments, the subject has necrozoospermia. In more particular embodiments, the subject has idiopathic necrozoospermia. 「 necrospermiaAs used herein, the term " " refers to a disorder characterized by a male subject's semen showing a low percentage of viable sperm in the ejaculate.

[0088] As used herein, the term "percentage of viable sperm" refers to the percentage of viable sperm relative to the total number of sperm in the ejaculate.

[0089] In a preferred embodiment, when a male patient has necrozoospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the male subject's semen show a percentage of viable sperm below the reference value. In a particular embodiment, the reference value is preferably the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition. The WHO has established the LRL for viable sperm in semen samples as 58% in said manual. Methods for determining the percentage of viable sperm in a male subject's semen or in a sample of the male subject's semen are well known to those skilled in the art. Non-limiting examples of such methods include the method of evaluating the amount of sperm with intact membranes in a sample described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, in particular the motility test disclosed in section 2.6 of said document.

[0090] In a particular embodiment, the subject has pyospermia. In a more particular embodiment, the subject has idiopathic pyospermia.

[0091] 「 pyospermia ( leukospermia 、 leucospermia 、 leukocytospermia 、 or pyospermiaThe term "aspermatospermia" as used herein refers to a condition characterized in that the semen of a male subject exhibits a concentration of white blood cells that exceeds the LRL. The term "concentration of white blood cells" as used herein refers to the total concentration of white blood cells per ejaculate divided by the volume of the ejaculate.

[0092] In a preferred embodiment, when a male patient has aspermatospermia, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject exhibit a concentration of white blood cells that exceeds the reference value. In a particular embodiment, the reference value is the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition. The WHO has established in the said manual the LRL for the concentration of white blood cells as 1×10 6 white blood cells per ml, when detected by a peroxidase test, i.e., established as peroxidase-positive white blood cells per ml. The said test is well-known to those skilled in the art and is described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition. Methods for determining the concentration of white blood cells in the semen of a male subject or in a semen sample of a male subject are well-known to those skilled in the art. Non-limiting examples of such methods include methods for determining the presence of white blood cells in a semen sample based on a peroxidase test, as described in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, in particular the method described in section 2.18 of the said document.

[0093] In a particular embodiment, the patient has a disorder characterized by a percentage of SDF that exceeds the reference value. In a more particular embodiment, the said disorder is a idiopathic disorder.

[0094] 「 sperm DNA fragmentation 」 or " SDFThe expression "」, when used in this specification, refers to DNA damage in sperm cells (from semen ejaculated during ejaculation or from testicular semen), i.e., single and / or double DNA breaks. Thus, Percentage of SDF or DNA fragmentation index (DFI) , when used in this specification, refers to the percentage of sperm cells showing DNA damage, i.e., single and / or double DNA breaks, relative to the total number of sperm in ejaculated semen. Having a certain percentage of SDF A male subject is understood as a subject characterized in that a sample of semen or ejaculated semen from the subject shows the percentage of said SDF. DNA repair occurs during sperm development, which is completed as the cessation of transcription and translation after sperm completion. As a result, sperm do not have a mechanism to repair DNA damage that occurs during their migration and storage in the epididymis or after ejaculation, which explains the sensitivity of sperm to DNA fragmentation and the presence of DNA fragmentation in the sperm of all male subjects. As will be understood by those skilled in the art, when the semen of a male subject has a high percentage of SDF, the ability of the semen of said subject to reach fertilization is low. In general terms, semen showing a percentage of DNA fragmentation of approximately 15% or less is considered to have good quality, i.e., a high ability to achieve fertilization. However, when the percentage of DNA fragmentation is above a percentage such as approximately 16% in some cases, the quality of the semen is considered low, i.e., the ability to achieve fertilization is low. Thus, a high percentage of SDF in a male subject is considered a disorder. When the cause of said disorder is unknown, this is considered an idiopathic disorder. " A semen sample shows x% SDF " or " A semen sample shows x% DNA fragmentation index (DFI)As used herein, the expression "x%" refers to a sample of semen in which x% of the sperm cells in the semen of the sample exhibit DNA fragmentation. Thus, as will be understood by those skilled in the art, a sample of semen showing a percentage of SDF of 15% or less has 15% or less of the sperm cells showing SDF and is generally considered a sample of semen of good quality, i.e., having a high ability to achieve fertilization. A sample of semen showing a percentage of SDF of 16% or more has 16% or more of the sperm cells showing SDF and is thus, in some cases, considered a sample of semen of low quality, i.e., having a low ability to achieve fertilization. In a preferred embodiment, if a male patient has a disorder characterized by a percentage of SDF above a reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a percentage of SDF above the reference value, where the reference value is preferably 16%. Similarly, if a male patient does not have a disorder characterized by a percentage of SDF above a reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a percentage of SDF below the reference value, where the reference value is preferably 16%.

[0095] Methods for determining the percentage of SDF or DFI in semen from or in a semen sample from a male subject are well known to those skilled in the art. Non-limiting examples of such methods include toluidine blue staining, CMA3 staining, acridine orange (AO) assay, SCSA, SCD test / Halo, comet assay, and the TUNEL assay described in Manesh Kumar Panner Selvam and Ashok Agarwal, 2018, Arab J Urol. 16(1):65-76.

[0096] In certain embodiments, the patient has a disorder characterized by a level of 8-OHdG in semen above a reference value, an 8-OHdG / 10 5 dG ratio, or a percentage of 8-OH-dG positive sperm cells. In more particular embodiments, the disorder is a idiopathic disorder. The term "8-hydroxy-2'-deoxyguanosine" or "8-OHdG" as used herein refers to an oxidized guanine base adduct formed when an OH radical damages DNA. This is considered a marker of DNA oxidation. Reactive oxygen species (ROS) can directly damage DNA and thus give rise to oxidized DNA adducts (e.g., 8-OHdG), which in turn destabilize the DNA structure and result in abasic sites that cause subsequent single-strand breaks. Thus, the presence of 8-OHdG can be associated with DNA oxidation and the subsequent appearance of DNA strand breaks. Indeed, DNA repair is limited in sperm and occurs only during specific stages of spermatogenesis. In particular, the repair machinery is no longer active during nuclear condensation in the epididymis, but sperm are exposed to oxidative damage in the epididymis and during transport in seminal plasma. DNA adducts induced by oxidative stress can be repaired by the oocyte, but single- or double-strand DNA breaks cannot be repaired. Thus, the presence of 8-OHdG in sperm cells can result in the appearance of DNA fragmentation and have a significant impact on fertilization and pregnancy outcomes. Thus, measuring the amount of 8-OH-dG in sperm cells in semen can determine the level of DNA damage in sperm cells in semen resulting from oxidative stress. Thus, in certain embodiments, in all aspects of the present invention, a disorder characterized by a level of 8-OH-dG in semen above a reference value, an 8-OH-dG / 10 5 dG ratio, or a percentage of 8-OH-dG positive sperm cells refers to a disorder characterized by a high level of DNA oxidation in semen. In another particular embodiment, in all aspects of the present invention, a level of 8-OH-dG in semen above a reference value, an 8-OH-dG / 10 5The disorder characterized by the dG ratio or the percentage of 8-OH-dG positive sperm cells refers to the disorder characterized by a high level of DNA damage in semen, preferably caused by oxidative stress in semen. In another embodiment, in all aspects of the present invention, the level of 8-OH-dG in semen exceeding the reference value, 8-OH-dG / 10 5 The disorder characterized by the dG ratio or the percentage of 8-OH-dG positive sperm cells refers to the disorder characterized by high oxidative stress in semen.

[0097] In a preferred embodiment, when a male patient has a condition characterized by the level of 8-OHdG in semen exceeding the reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a level of 8-OHdG exceeding the reference value. Similarly, when a male patient does not have a condition characterized by the level of 8-OHdG in semen exceeding the reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a level of 8-OH-dG below the reference value.

[0098] In another preferred embodiment, when a male patient has a condition characterized by the 8-OH-dG / 10 5 dG ratio in semen exceeding the reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show an 8-OH-dG / 10 5 dG ratio exceeding the reference value. Similarly, when a male patient has a condition characterized by the 8-OH-dG / 10 5In the case of not having a state characterized by the dG ratio, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject have an 8-OH-dG / 10 below the reference value 5 showing a dG ratio.

[0099] In another preferred embodiment, in the case where a male patient has a state characterized by the percentage of 8-OH-dG positive sperm cells in semen above the reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a percentage of 8-OH-dG positive sperm cells above the reference value. Similarly, in the case where a male patient does not have a state characterized by the percentage of 8-OH-dG positive sperm cells in semen above the reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a percentage of 8-OH-dG positive sperm cells below the reference value.

[0100] As used herein, the expression "level of 8-OH-dG in semen" refers to the amount of 8-OH-dG as defined above detected in semen. In certain embodiments, the level of 8-OH-dG in semen refers to the level of 8-OH-dG as defined above in ejaculated semen or in a semen sample. The level of 8-OH-dG in semen or in a semen sample can be measured by different methods. This is generally measured using immunofluorescence or a commercially available kit, such as the OxyDNA kit, followed by light microscopy, fluorescence microscopy or flow cytometry, preferably immunofluorescence followed by flow cytometry. Any of the said methods are well known to those skilled in the art and can be carried out as described in Vorilhon et al., 2018, Human reproduction, 33(4):553-562. In this case, the level of 8-OH-dG can be expressed as the mean fluorescence intensity (MIF) of the sample and can be measured in arbitrary units (a.u.). The reference values for the level of 8-OH-dG in a semen sample are 300, 325, 350, 375, 400, 425, 450, 475, 500, 510, 515, 520, 525, 530, 535, 540, 545, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 562, 565, 567, 570, 575, 580, 585, 590, 595, 600, 625, 650, 675, 700, 750, 800, 850, 900 arbitrary units, preferably at least 552 arbitrary units, when the method is based on immunofluorescence followed by flow cytometry as described in Vorilhon et al., 2018, Human reproduction, 33(4):553-562. As shown above, the level of 8-OH-dG in semen can refer to the level of 8-OH-dG in ejaculated semen or in a semen sample, and thus, in certain embodiments, the reference values for the parameters of the level of 8-OH-dG in semen are any of those described herein for the parameters of the level of 8-OH-dG in a semen sample.

[0101] "8-OH-dG / 10 5 dG ratio" as used herein, when referring to semen, refers to the amount of 8-OH-dG as defined above detected in semen divided by the amount of dG detected in the semen represented by the group of 10 5 , where dG refers to deoxyguanosine. In certain embodiments, the 8-OH-dG / 10 5 dG ratio refers to the 8-OH-dG / 10 5 dG ratio as defined in ejaculated semen or in a semen sample. The 8-OH-dG / 10 5 dG ratio can be measured by different methods. Non-limiting examples of such methods include high performance liquid chromatography with electrochemical detection (HPLC-EC), or gas chromatography-mass spectrometry (GC / MS). Such methods are well known to those skilled in the art and can be carried out, for example, according to the instructions found in Shen and Ong., 2000, Free Radical Biology and Medicine, 28(4):529-536. In this case, the reference value of the 8-OH-dG / 10 5 dG ratio in a semen sample is 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.25, 2.5, 2.75, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 80, 90, 100, preferably 1.5 8-OH-dG / 10 5 dG. As indicated above, the ratio of 8-OH-dG / 10 5 dG in semen can refer to the ratio of 8-OH-dG / 10 5 dG in ejaculated semen or in a semen sample, and thus, in certain embodiments, the reference value for the 8-OH-dG / 10 5 dG parameter in semen is any of those provided herein for the 8-OH-dG / 10 5 dG parameter in a semen sample.

[0102] "Percentage of 8-OH-dG positive sperm cells in semen", or "Percentage of 8-OH-dG positive sperm cells", or " Percentage of sperm cells having 8-OH-dGThe expression "___" when used in this specification refers to the percentage of sperm cells detected as positive for 8-OH-dG relative to the total number of sperm in semen. In certain embodiments, the percentage of 8-OH-dG positive sperm cells in semen refers to the percentage of 8-OH-dG as defined in ejaculated semen or in a semen sample. The percentage of 8-OH-dG positive sperm cells in semen can be measured by different methods. Non-limiting examples of such methods include immunofluorescence-based methods, or commercially available kits, such as the OxyDNA kit, followed by light microscopy, fluorescence microscopy or flow cytometry, preferably an immunofluorescence-based method followed by flow cytometry. Any of the said methods are well known to those skilled in the art and can be carried out as described in Vorilhon et al., 2018, Human reproduction, 33(4):553-562. In this case, the reference values for the percentage of 8-OH-dG positive sperm cells in a semen sample are 20%, 25%, 30%, 35%, 40%, 45%, 50%, 52%, 55%, 57%, 60%, 61%, 62%, 63%, 64%, 64.5%, 65%, 65.2%, 65.5%, 65.6%, 65.7%, 65.8%, 65.9%, 66%, 66.1%, 66.2%, 66.3%, 66.4%, 66.5%, 66.7%, 67%, 67.5%, 68%, 68.5%, 69%, 69.5%, 70%, 71%, 72%, 73%, 74%, 75%, 77%, 80%, 82%, 87.5%, 87%, 90%, 95%, 97%, 98%, 99% of 8-OH-dG positive sperm cells relative to the total number of sperm in the semen sample, preferably 65.8% of 8-OH-dG positive sperm cells relative to the total number of sperm in the semen sample. As shown above, the percentage of 8-OH-dG positive sperm cells in semen can refer to the percentage of 8-OH-dG positive sperm cells in ejaculated semen or in a semen sample, and thus, in certain embodiments, the reference values for the parameters of the percentage of 8-OH-dG positive sperm cells in semen are any of those described herein for the parameters of the percentage of 8-OH-dG positive sperm cells in a semen sample.

[0103] In certain embodiments, the patient has a disorder characterized by a static oxidation-reduction potential (sORP) that exceeds a reference value. In more particular embodiments, the disorder is a idiopathic disorder.

[0104] As used herein, the expression "static oxidation-reduction potential" or "sORP" refers to a measure of the balance between oxidants and reductants in a biological system that can be considered as a measure of oxidative stress in the sample. The expression "static oxidation-reduction potential in semen" or "sORP in semen" as used herein refers to the value of the oxidation-reduction potential as defined above when the biological system is semen. Thus, sORP in semen can be considered as a measure of the level of oxidative stress in semen. Thus, in certain embodiments, a disorder characterized by an sORP in semen that exceeds a reference value, as referred to in any aspect of the present invention, is a disorder characterized by high oxidative stress in semen.

[0105] In certain embodiments, the expression "sORP in semen" refers to the sORP in ejaculated semen or in a semen sample.

[0106] In a preferred embodiment, when a male patient has a disorder characterized by an sORP in semen that exceeds a reference value, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject exhibit an sORP that exceeds the reference value.

[0107] The sORP in semen or in a semen sample is directly related to the transfer of electrons from a reduced form to an oxidized form present in the sample of semen, or to the potential of the sample. Methods that enable measurement of said electron transfer or potential in semen or in a semen sample are well known to those skilled in the art. Non-limiting examples of methods that enable measurement of sORP in semen or in a semen sample include the MioXSYS system described in Agarwal A, Bui AD. Investig. Clin Urol. 2017;58(6):385-399. The value in mV provided by the type of said method is generally divided by the sperm concentration of the semen sample(s) being analyzed, such that sORP is generally provided as mV / 10 6 sperm / ml. In certain embodiments, reference values for sORP in a semen sample are 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1., 1.1, 1.2, 1.25, 1.27, 1.3, 1.31, 1.32, 1.33, 1.34, 1.35, 1.36, 1.37, 1.38, 1.39, 1.4, 1.41, 1.42, 1.43, 1.44, 1.45, 1.46, 1.47, 1.48, 1.49, 1.5, 1.51, 1.52, 1.53, 1.54, 1.55, 1.57, 1.6, 1.62, 1.65, 1.67, 1.7, 1.75, 1.8, 1.85, 1.9, 1.95, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.35, 5.6,5.7, 5.8, 5.9, 6.0, 6.25, 6.5, 6.75, 7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.75, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0 mV / 10 6 sperm / ml semen, preferably 1.34 mV / 10 6 sperm / ml semen, more preferably 1.36 mV / 10 6Spermatozoa / ml semen, and more preferably, 1.48 mV / 10 6 spermatozoa / ml semen. As indicated above, the sORP in semen can refer to the sORP in ejaculated semen or in a semen sample, and thus, in certain embodiments, the reference value for the parameter of sORP in semen is any of those described herein for the parameter of sORP in a semen sample.

[0108] In certain embodiments of the first aspect of the present invention, the total number and / or concentration of spermatozoa in a sample of semen of a male patient after treatment as referred to in the medical use of the present invention is above the reference value. Preferably, the total number and / or concentration of spermatozoa above the reference value is observed in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment as referred to in the medical use of the present invention.

[0109] In a preferred embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has oligospermia, and the total number and / or concentration of spermatozoa in a sample of semen of the patient after treatment as referred to in the medical use is above the reference value. Preferably, the total number of spermatozoa is above the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment as referred to in the medical use of the present invention. In another preferred embodiment, the concentration of spermatozoa is above the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment as referred to in the medical use of the present invention.

[0110] In one embodiment, the male patient is as defined in any of the embodiments of the first aspect of the present invention prior to the treatment referred to in the aspect of the present invention, and more preferably, the patient has oligospermia prior to the treatment, and the total number and / or concentration of sperm in the semen sample of the patient after the treatment is equal to or higher than the reference value. Preferably, the total number of sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples. In another preferred embodiment, the sperm concentration is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0111] The method for determining the number of sperm and the sperm concentration in a semen sample is provided above in the definition of "oligospermia".

[0112] 「 In a semen sample above the reference value, the Total number 」, when used herein, refers to the total number of sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0113] 「 In a semen sample equal to the reference value, the Total number 」, as used herein, refers to the total number of sperm in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001%, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1% higher than its reference value, where 100% represents the reference value. This also refers to the total number of sperm in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001%, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1% lower than its reference value, where 100% represents the reference value.

[0114] 「 In a semen sample below the reference value, the number of sperm 」, as used herein, refers to the total number of sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 90% lower than its reference value, where 100% corresponds to the reference value.

[0115] In certain embodiments, the reference value for the total number of sperm is 25×10 5 、30×10 5 、39×10 5 、1×10 6 、5×10 6 、10×10 6 、20×10 6 、25×10 6 、30×10 6 、39×10 6 、32×10 6 、35×10 6 、40×10 6 、45×10 6 、50×10 6 、60×10 6 、80×106 , 1×10 7 or 1×10 8 sperm cells per ejaculate, preferably 39×10 6 sperm cells per ejaculate. In another preferred embodiment, the reference value for the total number of sperm in a semen sample corresponds to the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, for the number of sperm per ejaculate, which is 39×10 6 sperm cells per ejaculate.

[0116] " In a semen sample above the reference value, the sperm concentration ", when used herein, refers to a sperm concentration in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0117] " In a semen sample equal to the reference value, the sperm concentration ", when used herein, refers to any sperm concentration in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, preferably less than 5%, more preferably less than 2%, and still more preferably less than 1% higher, where 100% represents the reference value. This also refers to any sperm concentration in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, preferably less than 5%, more preferably less than 2%, and still more preferably less than 1% lower, where 100% represents the reference value.

[0118] 「 In a semen sample below the reference value, the sperm concentration 」When used in this specification, the expression refers to the concentration of sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 90% lower than the reference value, where 100% corresponds to the reference value.

[0119] In certain embodiments, the reference value for the concentration of sperm in a semen sample is 5×10 5 、10×10 5 、15×10 5 、50×10 5 、1×10 6 、5×10 6 、10×10 6 、15×10 6 、20×10 6 、25×10 6 、50×10 6 、75×10 6 、10×10 7 、25×10 7 、50×10 7 、75×10 7 、or 10×10 8 sperm per 1 ml of semen, preferably 15×10 6 sperm per 1 ml of semen. In a preferred embodiment, the reference value for the concentration of semen in a semen sample corresponds to the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for the concentration of sperm per 1 ml of semen, which is 15×10 6 sperm per 1 ml of semen.

[0120] In another preferred embodiment, the reference value for the total number or concentration of sperm in a semen sample from a male subject is the total number or concentration of sperm in the semen sample of the male patient obtained prior to the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the total number or concentration of sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen from the male patient obtained prior to the commencement of the treatment referred to in the first aspect of the present invention. In one embodiment, where the reference value refers to the total number or concentration of sperm in more than 2 samples, the reference value corresponds to the average of the values of the parameter "total number of sperm" or "sperm concentration" obtained in each of said samples. In one embodiment, said samples are each samples obtained at different times prior to the commencement of the treatment referred to in the first aspect of the present invention.

[0121] In another specific embodiment, the reference value for the total number of sperm in a semen sample is less than 25×10 5 per ejaculate, less than 10×10 5 per ejaculate, less than 1×10 5 per ejaculate, less than 1×10 4 per ejaculate, less than 1×10 3 per ejaculate, less than 1×10 4 per ejaculate, less than 1×10 2 per ejaculate, less than 10 per ejaculate, preferably this is approximately 0 sperm per ejaculate, more preferably this is 0 sperm per ejaculate. In another specific embodiment, the reference value for the total number of sperm in a semen sample is the limit of quantification of sperm in the semen sample for the method of assessment used.

[0122] In another specific embodiment, since the reference value is the minimum for the parameter of "total number of sperm in a semen sample", the sperm in a semen sample below the reference value Total The expression "number" refers to the total number of sperm in a semen sample equal to the reference value. The expression "total number of sperm in a semen sample equal to the reference value" refers to the total number of sperm in a semen sample that is the reference value or is consistent with it. In another specific embodiment, the expression "total number of sperm in a semen sample exceeding the reference value" refers to any value higher than the reference value, preferably, this refers to an absolute value different from 0. In a preferred embodiment, this is at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 20, at least 100, at least 1×10 3 , at least 1×10 4 , at least 1×10 5 , at least 1×10 6 , at least 20×106 , at least 30×10 6 , at least 39×10 6 , at least 40×10 6 , at least 50×10 6 , at least 1×10 7 sperm per ejaculate, preferably, at least 39×10 6 sperm per ejaculate.

[0123] In another specific embodiment, the reference value for the sperm concentration in the semen sample is 5×10 5 less than, 4×10 5 less than, 1×10 5 less than, 1×10 4 less than, 1×10 3 less than, 1×10 2 less than, less than 10 sperm per 1 ml of semen, preferably, this is approximately 0 sperm per 1 ml of semen, more preferably, this is 0 sperm per 1 ml of semen. In another specific embodiment, the reference value for the sperm concentration in the semen sample is the limit of quantification of sperm in the semen sample for the evaluation method used. In this case, since the reference value is the minimum for the parameter of " Sperm in a semen sample concentration", " The expression "concentration of sperm in a semen sample below the reference value" refers to the concentration of sperm in a semen sample equal to the reference value . "Sperm in a semen sample equal to the reference value concentration " The expression refers to refers to the sperm concentration in the semen sample that is the reference value or is consistent with it. In another specific embodiment, "Concentration of sperm in a semen sample exceeding the reference value" the expression " refers to any value higher than the reference value, preferably, this refers to an absolute value different from 0. In a preferred embodiment, this is at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 20, at least 100, at least 1×10 3 , at least 1×10 4 , at least 1×10 5 , at least 1×10 6 , at least 1 5×10 6 、20×10 6 、25×10 6 、50×10 6 、75×10 6 、10×10 7 、50×10 7per milliliter of semen, preferably at least 15×10 6 refers to sperm per milliliter of semen.

[0124] In another specific embodiment, the patient of the first aspect of the present invention has azoospermia. The method for determining whether a subject has azoospermia is provided above in the definition of azoospermia. In another specific embodiment, all embodiments in which the term "oligospermia" is used are also applied herein by replacing the term "oligospermia" with "azoospermia". " Before the treatment referred to in the first aspect of the present invention " or " Before the start of the treatment referred to in the first aspect of the present invention " when used herein refers to any moment before treatment of the first aspect of the present invention. In a preferred embodiment, this is at least 1 hour before, at least 2 hours before, at least 5 hours before, at least 6 hours before, at least 8 hours before, at least 10 hours before, at least 12 hours before, at least 24 hours before, at least 1.5 days before, at least 2 days before, at least 4 days before, at least 5 days before, at least 6 days before, at least 1 week before, at least 1.5 weeks before, at least 2 weeks before, at least 3 weeks before, at least 1 month before, at least 2 months before, at least 3 months before, at least 4 months before, at least 5 months before, at least 6 months before, at least 1 year before, at least 1.5 years before, at least 2 years before, at least 5 years before, at least 10 years before, preferably at least 1 month before, more preferably at least 1 week before, even more preferably at least 1 day before, and still more preferably at least 1 hour before the start of said treatment.

[0125] " After the treatment referred to in the first aspect of the present invention " or " After the treatment referred to in the medical use of the present invention The expression "」" when used in this specification refers to any moment after the treatment of the first aspect of the present invention. In a preferred embodiment, this is the moment immediately after the end of the treatment, preferably at least 1 hour, at least 2 hours, at least 5 hours, at least 6 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, at least 1.5 days, at least 2 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 1.5 weeks, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 1 year, at least 1.5 years, at least 2 years, at least 5 years, at least 10 years after the end of the treatment, preferably at least 1 month, more preferably at least 1 week, even more preferably at least 1 day, and still more preferably at least 1 hour after the end of the treatment.

[0126] In a specific embodiment of the first aspect of the present invention, the percentage of progressively motile sperm in a sample of semen from a male patient after the treatment referred to in the medical use of the present invention is equal to or higher than a reference value. Preferably, the percentage of progressively motile sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen from a male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0127] In a preferred embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has asthenospermia, and the percentage of progressively motile sperm in a sample of the semen of the male patient after the treatment referred to in the medical use of the present invention is equal to or higher than a reference value. Preferably, the percentage of progressively motile sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0128] In an embodiment, the male patient is as defined in any of the embodiments of the first aspect of the present invention before the treatment referred to in the above aspect of the present invention. Preferably, the patient has asthenospermia before the treatment, and the percentage of progressively motile sperm in a sample of the semen of the male patient after the treatment is equal to or higher than a reference value. Preferably, the percentage of progressively motile sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0129] The expression "progressive motility" is defined above. The method for determining the percentage of progressively motile sperm in a semen sample is provided above in the definition of asthenospermia.

[0130] 「 Percentage of progressively motile sperm in a semen sample exceeding the reference value The expression "」, when used in this specification, refers to any percentage of progressively motile sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, where 100% corresponds to the reference value.

[0131] 「 Percentage of progressively motile sperm in a semen sample equal to the reference value The expression "」, when used in this specification, refers to any percentage of progressively motile sperm in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, where 100% represents the reference value. This also refers to any percentage of progressively motile sperm in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, where 100% represents the reference value.

[0132] 「 Percentage of progressively motile sperm in a semen sample below the reference value The expression "」, when used in this specification, refers to the percentage of progressively motile sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than the reference value, where 100% corresponds to the reference value.

[0133] In certain embodiments, the reference value for the percentage of progressively motile sperm in a semen sample is 10%, 15%, 20%, 25%, 30%, 31%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%, 34.5%, 35%, 35.5%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 47%, 50%, 52%, 55%, 60%, 65%, 70%, 80%, 90%, 95%, preferably 30%, more preferably 32%. In a preferred embodiment, the reference value for the percentage of progressively motile sperm in a semen sample is the LRL established by the WHO in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th edition, for the percentage of progressively motile sperm in a semen sample, which is 32%.

[0134] In another preferred embodiment, the reference value for the percentage of progressively motile sperm in a semen sample of a male subject is the percentage of progressively motile sperm in a sample of semen of said male patient obtained prior to the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the percentage of progressively motile sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, more preferably at least 2 samples of semen of said male patient obtained prior to the commencement of the treatment referred to in the first aspect of the present invention. In one embodiment, where the reference value refers to the percentage of progressively motile sperm in two or more samples obtained prior to the commencement of the treatment referred to in the first aspect of the present invention, said reference value corresponds to the average of the values of the parameter "percentage of progressively motile sperm" obtained in each of said samples. In one embodiment, said samples are each samples obtained at different times prior to the commencement of the treatment referred to in the first aspect of the present invention.

[0135] The terms "post-treatment" and "pre-treatment" are already defined.

[0136] In a specific embodiment of the first aspect of the present invention, the percentage of morphologically normal sperm in a sample of semen from a male patient after the treatment referred to in the medical use of the present invention is equal to or higher than a reference value. Preferably, the percentage of morphologically normal sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen from a male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0137] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has teratozoospermia, and the percentage of morphologically normal sperm in a sample of semen from the patient after the treatment referred to in the medical use of the present invention is equal to or higher than a reference value. Preferably, the percentage of morphologically normal sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen from a male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0138] In an embodiment, the male patient is as defined in any of the embodiments of the first aspect of the present invention before the treatment referred to in the above aspect of the present invention. More preferably, the patient has teratozoospermia before the treatment, and the percentage of morphologically normal sperm in a sample of semen from the male patient after the treatment is equal to or higher than a reference value. Preferably, the percentage of morphologically normal sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen from a male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0139] 「 Morphologically normal sperm 」 The expression is provided above. The method for determining the percentage of morphologically normal sperm in a sample is provided above in the definition of teratospermia.

[0140] 「 Percentage of morphologically normal sperm in a semen sample exceeding the reference value 」 When used in this specification, the expression refers to any percentage of morphologically normal sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0141] 「 Percentage of morphologically normal sperm in a semen sample equal to the reference value 」 When used in this specification, the expression refers to any percentage of morphologically normal sperm that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, or less than 0.001% higher than its reference value, where 100% represents the reference value. This also refers to any percentage of morphologically normal sperm that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, or less than 0.001% lower than its reference value, where 100% represents the reference value.

[0142] 「 Percentage of morphologically normal sperm in a semen sample below the reference value The expression "」" when used in this specification refers to the percentage of morphologically normal sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, preferably at least 25%, more preferably at least 50%, even more preferably at least 100% lower than the reference value, where 100% corresponds to the reference value.

[0143] In certain embodiments, the reference value for the percentage of morphologically normal sperm in a sample is 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 8%, 9%, 10%, 12%, 15%, 17%, 20%, 22%, 25%, 30%, 35%, 40%, preferably 4%. In a preferred embodiment, the reference value for the percentage of morphologically normal sperm in a semen sample is the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, which is 4%.

[0144] In another preferred embodiment, the reference value for the percentage of morphologically normal sperm in a semen sample of a male subject is the percentage of morphologically normal sperm in the semen sample of the male patient obtained before the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the percentage of morphologically normal sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male patient obtained before the treatment referred to in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the percentage of morphologically normal sperm in two or more samples obtained before the start of the treatment referred to in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "percentage of morphologically normal sperm in the semen sample" obtained in each of the samples. In another embodiment, the samples are each samples obtained at different times before the treatment referred to in the first aspect of the present invention is initiated.

[0145] The terms "post-treatment" and "pre-treatment" are already defined.

[0146] In a specific embodiment of the first aspect of the present invention, the volume of semen per ejaculate in a semen sample of a male patient after the treatment referred to in the medical use of the present invention is equal to or greater than the reference value. As indicated in the definition of "semen sample", a semen sample of a male subject preferably contains or consists of the semen of an ejaculate from the subject. Thus, as will be understood by those skilled in the art, the volume of semen per ejaculate in a semen sample is the volume of the semen sample. Also, as will be understood by those skilled in the art, the volume of semen per ejaculate in a semen sample is the volume of the ejaculate contained in or consisting of the sample.

[0147] In the case where a sample of semen of interest contains two or more ejaculates of the subject, the volume of semen per ejaculate in the semen sample is the volume of one of the ejaculates. The volume can be readily determined by one skilled in the art by dividing the volume of the sample by the number of ejaculates contained in the sample or of which the semen sample consists.

[0148] Preferably, the volume of semen per ejaculate in a sample of semen of a male patient is above a reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples, of semen of the male patient after treatment as referred to in the medical use of the present invention.

[0149] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has oligospermia, and the volume of semen per ejaculate in a sample of semen of the male patient after treatment as referred to in the medical use of the present invention is above a reference value. Preferably, the volume of semen per ejaculate in the semen sample is above a reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples, of semen of the male patient after treatment as referred to in the medical use of the present invention.

[0150] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention before the treatment referred to in the above aspect of the present invention. More preferably, the patient has oligospermia before the treatment, and the volume of semen per ejaculate in the semen sample of the male patient after the treatment referred to in the medical use of the present invention is equal to or greater than the reference value. Preferably, the volume of semen per ejaculate is equal to or greater than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0151] The method for determining the volume of semen in the ejaculate of a male patient is provided above in the definition of oligospermia.

[0152] 「 Volume of semen per ejaculate in a semen sample below the reference value 」 When used in this specification, this expression refers to a volume of semen per ejaculate that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than the reference value, where 100% corresponds to the reference value.

[0153] 「 Volume of semen per ejaculate in semen samples above the reference value 」 When used in this specification, this expression refers to a volume of semen per ejaculate that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150%, where 100% corresponds to the reference value.

[0154] The expression " Volume of semen per ejaculate in semen samples equal to the reference value ", as used herein, refers to the volume of semen per ejaculate that is equal to, or substantially equal to, the reference value for the volume of semen in the ejaculate. Thus, the volume of semen per ejaculate is equal to the reference value where it is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than the reference value, where 100% represents the reference value. This also applies where it is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than the reference value, where 100% represents the reference value.

[0155] In another preferred embodiment, the reference value for the volume of semen per ejaculate is 5 ml, 4.5 ml, 4 ml, 3.5 ml, 3 ml, 2.5 ml, 2 ml, 1.5 ml, 1 ml, 0.5 ml, 0.25 ml, preferably 1.5 ml. In a preferred embodiment, the reference value corresponds to the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, and is 1.5 ml.

[0156] In another preferred embodiment, the reference value for the volume of semen per ejaculate in a semen sample is the volume of semen per ejaculate of the male subject obtained prior to the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the volume of semen per ejaculate in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of the male patient obtained prior to the start of the treatment referred to in the first aspect of the present invention.

[0157] In one embodiment, when the reference value refers to the volume of semen per ejaculate in two or more samples obtained before the start of the treatment referred to in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "volume of semen per ejaculate in the semen sample" obtained in each of the samples. In one embodiment, the samples are each sample obtained at different times before the treatment referred to in the first aspect of the present invention is initiated.

[0158] In another specific embodiment, the reference value for the volume of semen per ejaculate in the semen sample is lower than 0.25 ml, lower than 0.2 ml, lower than 0.1 ml, lower than 0.05 ml, lower than 0.01 ml, lower than 0.005 ml, preferably, this is approximately 0 ml, and more preferably, this is 0 ml.

[0159] In another specific embodiment, since the reference value is the minimum for the parameter "volume of semen per ejaculate in the semen sample", the expression "volume of semen per ejaculate in a semen sample below the reference value" refers to the volume of semen per ejaculate in a semen sample equal to the reference value. The expression "volume of semen per ejaculate in a semen sample equal to the reference value" refers to the volume of semen per ejaculate in a semen sample that is the reference value or coincides with it. In another specific embodiment, the expression "volume of semen per ejaculate in a semen sample above the reference value" refers to any value higher than the reference value, preferably, this refers to an absolute value different from 0. In a preferred embodiment, this refers to the volume of semen per ejaculate in a semen sample of at least 0.001 ml, at least 0.005 ml, at least 0.01 ml, at least 0.1 ml, at least 0.2 ml, at least 0.5 ml, at least 1 ml, at least 1.5 ml, at least 1.75 ml, at least 2 ml, at least 2.5 ml, preferably, at least 1.5 ml.

[0160] In another specific embodiment, the patient of the first aspect of the present invention has azoospermia. The method for determining whether a subject has azoospermia is provided in the above definition of azoospermia. In another specific embodiment, all embodiments in which the term "oligospermia" is used are also applied herein by replacing the term "oligospermia" with "azoospermia".

[0161] The terms "post-treatment" and "pre-treatment" have already been defined.

[0162] In a specific embodiment of the first aspect of the present invention, the percentage of SDF in a sample of semen of a male patient after treatment referred to in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the percentage of SDF is any of those defined below for the parameter of the percentage of SDF. Preferably, the percentage of SDF is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment referred to in the medical use of the present invention.

[0163] In a preferred embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has a disorder characterized by a percentage of SDF above a reference value, and the percentage of SDF in a sample of semen of the male patient after treatment mentioned in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the percentage of SDF is any of those defined below for the parameter of the percentage of SDF. Preferably, the percentage of SDF is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of the male patient after treatment mentioned in the medical use of the present invention.

[0164] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention as mentioned in the foregoing aspect of the present invention. More preferably, the patient has a disorder characterized by a percentage of SDF above a reference value before the treatment mentioned in the foregoing aspect, and the percentage of SDF in a sample of semen of the male patient after the treatment is below the reference value. In a preferred embodiment, the reference value for the percentage of SDF is any of those defined below for the parameter of the percentage of SDF. In another preferred embodiment, the percentage of SDF is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of the male patient after treatment mentioned in the medical use of the present invention. The method for determining the percentage of SDF is provided above in the definition of sperm DNA fragmentation.

[0165] 「 Percentage of SDF in semen samples above the reference valueThe expression "」, when used in this specification, refers to the percentage of SDF in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, where 100% corresponds to the reference value.

[0166] 「 Percentage of SDF in semen samples equal to the reference value The expression "」, when used in this specification, refers to any percentage of SDF in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, where 100% represents the reference value. This also refers to the percentage of SDF that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, where 100% represents the reference value.

[0167] 「 Percentage of SDF in semen samples below the reference value The expression "」, when used in this specification, refers to the percentage of SDF in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than the reference value, where 100% corresponds to the reference value.

[0168] In a preferred embodiment, any reference value mentioned above for the percentage of SDF in a semen sample is the percentage of SDF associated with a low or poor quality of the semen sample. Thus, the reference value is 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 23%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 42%, 45%, 47%, 50%, preferably 30%, more preferably 20%, even more preferably 18%, still more preferably 16%, and even still more preferably 15%.

[0169] In another preferred embodiment, the reference value for the percentage of SDF in a semen sample of a male subject is the percentage of SDF in the sample of semen of the male patient obtained prior to the treatment mentioned in the first aspect of the present invention. In another preferred embodiment, this refers to the percentage of SDF in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of the male patient obtained before the treatment mentioned in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the percentage of SDF in two or more samples obtained before the start of the treatment mentioned in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "percentage of SDF in a semen sample" obtained in each of the samples. In one embodiment, the samples are each sample obtained at different times before the treatment mentioned in the first aspect of the present invention is initiated.

[0170] In another embodiment, the reference value for the percentage of SDF in a semen sample of a male subject is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of male subjects considered to be healthy, preferably male subjects considered to be fertile. In another embodiment, the reference value for the percentage of SDF in a semen sample of a male subject is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of different male subjects considered to be healthy, preferably different male subjects considered to be fertile. The definition of the term "fertile" or "fertility" is provided in the second aspect of the present invention and applies to this aspect of the present invention.

[0171] The terms "post-treatment" and "pre-treatment" are already defined.

[0172] In a specific embodiment of the first aspect of the present invention, the level of 8-OH-dG in a sample of semen of a male patient after treatment referred to in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the level of 8-OH-dG in a semen sample is any of those defined below for the parameters of the level of 8-OH-dG in a semen sample. Preferably, the level of 8-OH-dG in a semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of male patients after treatment referred to in the medical use of the present invention.

[0173] In a preferred embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has a disorder characterized by a level of 8-OH-dG in semen exceeding a reference value, and the level of 8-OH-dG in a sample of semen of the male patient after treatment referred to in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the level of 8-OH-dG in the semen sample is any of those defined below for the parameter of the level of 8-OH-dG in semen. Preferably, the level of 8-OH-dG in the semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen of the male patient after treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0174] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention before the treatment mentioned in the above aspect of the present invention. More preferably, the patient has a disorder characterized by a level of 8-OH-dG in semen exceeding a reference value before the treatment, and the level of 8-OH-dG in a semen sample of the male patient after the treatment is below the reference value. In a preferred embodiment, the reference value for the level of 8-OH-dG in a semen sample is any of those defined below for the parameters of the level of 8-OH-dG in a semen sample. In another preferred embodiment, the level of 8-OH-dG in a semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of the semen of the male patient after the treatment mentioned in the medical use of the present invention. The method for determining the level of 8-OH-dG in semen or in a semen sample is provided above in the definition of "level of 8-OH-dG in semen".

[0175] 「 In semen samples above the reference value The expression "level of 8-OH-dG", when used herein, refers to a level of 8-OH-dG in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0176] 「 In semen samples equal to the reference valueAs used herein, the expression "level of 8-OH-dG" refers to any percentage of SDF in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1%, where 100% represents the reference value. This also refers to a level of 8-OH-dG that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1%, where 100% represents the reference value.

[0177] “ In semen samples below the reference value As used herein, the expression "level of 8-OH-dG" refers to a level of 8-OH-dG in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than its reference value, preferably at least 25%, more preferably at least 50%, and even more preferably at least 90%, where 100% corresponds to the reference value.

[0178] In a preferred embodiment, In semen samplesFor the level of 8-OH-dG, any reference value mentioned above is 300, 325, 350, 375, 400, 425, 450, 475, 500, 510, 515, 520, 525, 530, 535, 540, 545, 550, 551, 552, 553, 554, 555, 556, 557, 558, 559, 560, 562, 565, 567, 570, 575, 580, 585, 590, 595, 600, 625, 650, 675, 700, 750, 800, 850, 900 arbitrary units, preferably at least 552 arbitrary units, when the method is based on immunofluorescence followed by flow cytometry as described in Vorilhon et al., 2018, Human reproduction, 33(4):553-562.

[0179] In another preferred embodiment, the reference value for the level of 8-OH-dG in a semen sample of a male subject is the level of 8-OH-dG in the semen sample of the male patient obtained before the treatment mentioned in the first aspect of the present invention. In another preferred embodiment, this refers to the level of 8-OH-dG in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male patient obtained before the treatment mentioned in the first aspect of the present invention is initiated. In an embodiment, when the reference value refers to the level of 8-OH-dG in two or more samples obtained before the start of the treatment mentioned in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "level of 8-OH-dG in the semen sample" obtained in each of the samples. In an embodiment, the samples are each sample obtained at different time points before the treatment mentioned in the first aspect of the present invention is initiated.

[0180] In another embodiment, the reference value for the level of 8-OH-dG in a semen sample of a male subject is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of male subjects considered to be healthy, preferably male subjects considered to be fertile. In another embodiment, the reference value for the level of 8-OH-dG in a semen sample of a male subject is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of different male subjects considered to be healthy, preferably different male subjects considered to be fertile. The definition of the term "fertile" or "fertility" is provided in the second aspect of the present invention and is applicable to this aspect of the present invention.

[0181] In another specific embodiment, as shown above, the percentage of 8-OH-dG positive sperm cells in semen refers to the percentage of 8-OH-dG positive sperm cells in ejaculated semen or in a semen sample. Thus, as will be understood by those skilled in the art, the reference value for the parameter of "level of 8-OH-dG in semen" is interchangeable with the reference value for the parameter of "level of 8-OH-dG in a semen sample". In an embodiment, the reference value for the parameter of "level of 8-OH-dG in semen" is any of those provided above for the parameter of "level of 8-OH-dG in a semen sample".

[0182] The terms "post-treatment" and "pre-treatment" have already been defined.

[0183] In a specific embodiment of the first aspect of the present invention, 8-OH-dG / 10 in semen in a sample of semen of a male patient after treatment as referred to in the medical use of the present invention 5The 8-OH-dG / 10 dG ratio is below the reference value. In a preferred embodiment, the 8-OH-dG / 10 5 The reference value for the 8-OH-dG / 10 dG ratio is the 8-OH-dG / 10 5 The parameter of the 8-OH-dG / 10 dG ratio is any of those defined below. Preferably, the 8-OH-dG / 10 5 The 8-OH-dG / 10 dG ratio is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of a male patient after the treatment mentioned in the medical use of the present invention, preferably in at least 2 samples.

[0184] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has a disorder characterized by an 8-OH-dG / 10 5 dG ratio in semen above the reference value, and the 8-OH-dG / 10 5 dG ratio in the sample of the semen of the male patient after the treatment mentioned in the medical use of the present invention is below the reference value. In a preferred embodiment, the 8-OH-dG / 10 5 The reference value for the 8-OH-dG / 10 dG ratio is the 8-OH-dG / 10 5 The parameter of the 8-OH-dG / 10 dG ratio is any of those defined below. Preferably, the 8-OH-dG / 10 5 The 8-OH-dG / 10 dG ratio is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of a male patient after the treatment mentioned in the medical use of the present invention, preferably in at least 2 samples.

[0185] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention before the treatment mentioned in the above aspect of the present invention, and more preferably, the patient has a disorder characterized by an 8-OH-dG / 10 5 dG ratio in semen exceeding a reference value before the treatment, and the 8-OH-dG / 10 5 dG ratio in the semen sample of the male patient after the treatment is below the reference value. In a preferred embodiment, the reference value for the 8-OH-dG / 10 5 dG ratio is any of those defined below for the parameter of the 8-OH-dG / 10 5 dG ratio in the semen sample. In another preferred embodiment, the 8-OH-dG / 10 5 dG ratio is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment mentioned in the medical use of the present invention, preferably in at least 2 samples. The method for determining the 8-OH-dG / 10 5 dG ratio in semen or in a semen sample is provided above in the definition of "8-OH-dG / 10 5 dG ratio in semen".

[0186] " In semen samples above the reference value 8-OH-dG / 10 5 dG ratio", when used herein, is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher than the reference value, for the 8-OH-dG / 105 Refers to the dG ratio, where 100% corresponds to the reference value.

[0187] 「 In semen samples equal to the reference value 8-OH-dG / 10 5 When the expression "dG ratio" is used in this specification, it refers to any percentage of SDF in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than the reference value, where 100% represents the reference value. This also refers to 8-OH-dG / 10 that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than the reference value, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1% lower. 5 Refers to the dG ratio, where 100% represents the reference value.

[0188] 「 In semen samples below the reference value 8-OH-dG / 10 5 When the expression "dG ratio" is used in this specification, it refers to 8-OH-dG / 10 in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than the reference value, preferably at least 25%, more preferably at least 50%, and even more preferably at least 90% lower. 5 Refers to the dG ratio, where 100% corresponds to the reference value.

[0189] In a preferred embodiment, In semen samples 8-OH-dG / 10 5Any reference value mentioned above for the 8-OH-dG / 10 dG ratio is 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.25, 2.5, 2.75, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 80, 90, 100, preferably 1.5 8-OH-dG / 10 5 dG.

[0190] In another preferred embodiment, the reference value for the 8-OH-dG / 10 dG ratio in a semen sample from a male subject is the 8-OH-dG / 10 5 dG ratio in a sample of semen from said male patient obtained prior to the treatment mentioned in the first aspect of the present invention. In another preferred embodiment, this is the 8-OH-dG / 10 5 dG ratio in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen from said male patient obtained prior to the commencement of the treatment mentioned in the first aspect of the present invention. 5 In one embodiment, where the reference value refers to the 8-OH-dG / 10 5 dG ratio in two or more samples obtained prior to the commencement of the treatment mentioned in the first aspect of the present invention, said reference value corresponds to the mean of the values of the parameter "8-OH-dG / 10 5 dG ratio in semen sample" obtained in each of said samples. In one embodiment, said samples are each samples obtained at different times prior to the commencement of the treatment mentioned in the first aspect of the present invention.

[0191] In another embodiment, the 8-OH-dG / 10 5The reference value for the 8-OH-dG / 10dG ratio is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen from male subjects considered to be healthy, preferably male subjects considered to be fertile. 5 It is the 8-OH-dG / 10dG ratio. In another embodiment, the 8-OH-dG / 10 5 The reference value for the 8-OH-dG / 10dG ratio is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen from different male subjects considered to be healthy, preferably different male subjects considered to be fertile. 5 It is the 8-OH-dG / 10dG ratio. The definition of the term "fertile" or "fertility" is provided in the second aspect of the present invention and applies to this aspect of the present invention.

[0192] In another specific embodiment, as shown above, the 8-OH-dG / 10 5 dG ratio refers to the 8-OH-dG / 10 5 dG ratio in ejaculated semen or in a semen sample. Thus, as will be understood by those skilled in the art, the reference value for the parameter "8-OH-dG / 10 5 dG ratio in semen" is compatible with the reference value for the parameter "8-OH-dG / 10 5 dG ratio in a semen sample". In an embodiment, the reference value for the parameter "8-OH-dG / 10 5 dG ratio in semen" is any of those provided above for the parameter "8-OH-dG / 10 5 dG ratio in a semen sample".

[0193] The terms "after treatment" and "before treatment" are already defined.

[0194] In a specific embodiment of the first aspect of the present invention, the percentage of 8-OH-dG positive sperm cells in semen in a semen sample of a male patient after treatment mentioned in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in the semen sample is any of those defined below for the parameter of the percentage of 8-OH-dG positive sperm cells in the semen sample. Preferably, the percentage of 8-OH-dG positive sperm cells in the semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen of a male patient after treatment mentioned in the medical use of the present invention, preferably in at least 2 samples.

[0195] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has a disorder characterized by a percentage of 8-OH-dG positive sperm cells in semen exceeding the reference value, and the percentage of 8-OH-dG positive sperm cells in a semen sample of a male patient after treatment mentioned in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in the semen sample is any of those defined below for the parameter of the percentage of 8-OH-dG positive sperm cells in the semen sample. Preferably, the percentage of 8-OH-dG positive sperm cells in the semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen of a male patient after treatment mentioned in the medical use of the present invention, preferably in at least 2 samples.

[0196] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention before the treatment referred to in the above aspect of the present invention. More preferably, the patient has a disorder characterized by a percentage of 8-OH-dG positive sperm cells in semen exceeding a reference value before the treatment, and the percentage of 8-OH-dG positive sperm cells in a semen sample of the male patient after the treatment is below the reference value. In a preferred embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in a semen sample is any of those defined below for the parameter of the percentage of 8-OH-dG positive sperm cells in a semen sample. In another preferred embodiment, the percentage of 8-OH-dG positive sperm cells in a semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples. The method for determining the percentage of 8-OH-dG positive sperm cells in semen or in a semen sample is provided above in the definition of "percentage of 8-OH-dG positive sperm cells in semen".

[0197] 「 In semen samples above the reference value The expression "percentage of 8-OH-dG positive sperm cells", as used herein, refers to a percentage of 8-OH-dG positive sperm cells in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0198] 「 In semen samples equal to the reference value When the expression "percentage of 8-OH-dG positive sperm cells" is used herein, it refers to any percentage of SDF in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, where 100% represents the reference value. This also refers to a percentage of 8-OH-dG positive sperm cells that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, where 100% represents the reference value.

[0199] 「 In semen samples below the reference value When the expression "percentage of 8-OH-dG positive sperm cells" is used herein, it refers to a percentage of 8-OH-dG positive sperm cells in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than its reference value, where 100% corresponds to the reference value.

[0200] In a preferred embodiment, In semen samplesFor the percentage of 8-OH-dG positive sperm cells, any of the reference values mentioned above are 20%, 25%, 30%, 35%, 40%, 45%, 50%, 52%, 55%, 57%, 60%, 61%, 62%, 63%, 64%, 64.5%, 65%, 65.2%, 65.5%, 65.6%, 65.7%, 65.8%, 65.9%, 66%, 66.1%, 66.2%, 66.3%, 66.4%, 66.5%, 66.7%, 67%, 67.5%, 68%, 68.5%, 69%, 69.5%, 70%, 71%, 72%, 73%, 74%, 75%, 77%, 80%, 82%, 87.5%, 87%, 90%, 95%, 97%, 98%, 99% of the total number of sperm in the semen sample, preferably 65.8% of the total number of sperm in the semen sample.

[0201] In another preferred embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in a semen sample of a male subject is the percentage of 8-OH-dG positive sperm cells in the semen sample of the male patient obtained prior to the treatment mentioned in the first aspect of the present invention. In another preferred embodiment, this refers to the percentage of 8-OH-dG positive sperm cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male patient obtained before the treatment mentioned in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the percentage of 8-OH-dG positive sperm cells in two or more samples obtained before the start of the treatment mentioned in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "percentage of 8-OH-dG positive sperm cells in the semen sample" obtained in each of the samples. In one embodiment, the samples are each sample obtained at different time points before the treatment mentioned in the first aspect of the present invention is initiated.

[0202] In another embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in a semen sample from a male subject is the percentage of 8-OH-dG positive sperm cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples from male subjects considered to be healthy, preferably male subjects considered to be fertile. In another embodiment, the reference value for the percentage of 8-OH-dG positive sperm cells in a semen sample from a male subject is the percentage of 8-OH-dG positive sperm cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples from different male subjects considered to be healthy, preferably different male subjects considered to be fertile. The definition of the term "fertile" or "fertility" is provided in the second aspect of the present invention and applies to this aspect of the present invention.

[0203] In another specific embodiment, as indicated above, the percentage of 8-OH-dG positive sperm cells in semen refers to the percentage of 8-OH-dG positive sperm cells in ejaculated semen or in a semen sample. Thus, as will be understood by those skilled in the art, the reference value for the parameter "percentage of 8-OH-dG positive sperm cells in semen" is interchangeable with the reference value for the parameter "percentage of 8-OH-dG positive sperm cells in a semen sample". In an embodiment, the reference value for the parameter "percentage of 8-OH-dG positive sperm cells in semen" is any of those provided above for the parameter "percentage of 8-OH-dG positive sperm cells in a semen sample".

[0204] The terms "post-treatment" and "pre-treatment" are already defined.

[0205] In a particular embodiment of the first aspect of the present invention, the sORP in a sample of semen from a male patient after the treatment referred to in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for sORP in the semen sample is any of those defined below for the parameters of sORP in the semen sample. Preferably, the sORP in the semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen from a male patient after the treatment referred to in the medical use of the present invention.

[0206] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has a disorder characterized by an sORP in semen above the reference value, and the sORP in a sample of semen from a male patient after the treatment referred to in the medical use of the present invention is below the reference value. In a preferred embodiment, the reference value for sORP in the semen sample is any of those defined below for the parameters of sORP in the semen. Preferably, the sORP in the semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen from a male patient after the treatment referred to in the medical use of the present invention.

[0207] In one embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention prior to the treatment mentioned in the above aspect of the present invention. Preferably, the patient has a disorder characterized by sORP in semen exceeding a reference value prior to the treatment, and the sORP in a semen sample of the male patient after the treatment is below the reference value. In a preferred embodiment, the reference value for sORP in a semen sample is any of those defined below for the parameters of sORP in a semen sample. In another preferred embodiment, the sORP in a semen sample is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of semen of the male patient after the treatment mentioned in the medical use of the present invention, preferably in at least 2 samples. The method for determining sORP in semen or in a semen sample is provided above in the definition of "sORP in semen".

[0208] 「 In semen samples above the reference value The expression "sORP" as used herein refers to sORP in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0209] 「 In semen samples equal to the reference valueThe expression "sORP", as used herein, refers to any percentage of sORP in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001%, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1% higher than its reference value, where 100% represents the reference value. This also refers to a level of sORP that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001%, preferably less than 5%, more preferably less than 2%, and even more preferably less than 1% lower than its reference value, where 100% represents the reference value.

[0210] 「 sORP in semen samples below the reference value The expression "", as used herein, refers to sORP in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 90% lower than its reference value, where 100% corresponds to the reference value.

[0211] In a preferred embodiment, any of the reference values mentioned above for sORP in a semen sample is 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1., 1.1, 1.2, 1.25, 1.27, 1.3, 1.31, 1.32, 1.33, 1.34, 1.35, 1.36, 1.37, 1.38, 1.39, 1.4, 1.41, 1.42, 1.43, 1.44, 1.45, 1.46, 1.47, 1.48, 1.49, 1.5, 1.51, 1.52, 1.53, 1.54, 1.55, 1.57, 1.6, 1.62, 1.65, 1.67, 1.7, 1.75, 1.8, 1.85, 1.9, 1.95, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.35, 5.6, 5.7, 5.8, 5.9, 6.0, 6.25, 6.5, 6.75, 7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.75, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0 mV / 10 6 spermatozoa / ml semen, preferably, 1.34 mV / 10 6 spermatozoa / ml semen, more preferably, 1.36 mV / 10 6 spermatozoa / ml semen, even more preferably, 1.48 mV / 10 6 spermatozoa / ml semen.

[0212] In another preferred embodiment, the reference value for sORP in a semen sample of a male subject is the sORP in a sample of semen of the male patient obtained before the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the sORP in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of the male patient obtained before the treatment referred to in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the sORP in two or more samples obtained before the start of the treatment referred to in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "sORP in semen sample" obtained in each of the samples. In one embodiment, the samples are each sample obtained at different times before the treatment referred to in the first aspect of the present invention is initiated.

[0213] In another embodiment, the reference value for sORP in a semen sample of a male subject is the sORP in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of a male subject considered to be healthy, preferably a male subject considered to be fertile. In another embodiment, the reference value for sORP in a semen sample of a male subject is the sORP in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of semen of different male subjects considered to be healthy, preferably different male subjects considered to be fertile. The definition of the term "fertile" or "fertility" is provided in the second aspect of the present invention and applies to this aspect of the present invention.

[0214] In another specific embodiment, as shown above, sORP in semen refers to sORP in ejaculated semen or in a semen sample. Thus, as understood by those skilled in the art, the reference value for the parameter of "sORP in semen" is interchangeable with the reference value for the parameter of "sORP in a semen sample". In an embodiment, the reference value for the parameter of "sORP in semen" is any of those provided above for the parameter of "sORP in a semen sample".

[0215] The terms "after treatment" and "before treatment" are already defined.

[0216] In a specific embodiment of the first aspect of the present invention, the number of viable sperm in a sample of semen of a male patient after treatment referred to in the medical use of the present invention is equal to or greater than the reference value. Preferably, the percentage of viable sperm is equal to or greater than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment referred to in the medical use of the present invention.

[0217] In a preferred embodiment, the male patient in the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has necrospermia, and the percentage of viable sperm in a sample of semen of the said patient after treatment referred to in the medical use of the present invention is equal to or greater than the reference value. Preferably, the percentage of viable sperm is equal to or greater than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of semen of a male patient after treatment referred to in the medical use of the present invention.

[0218] In one embodiment, the male patient is as defined in any of the embodiments of the first aspect of the present invention prior to the treatment referred to in the aspect of the present invention. More preferably, the patient has asthenozoospermia prior to the treatment, and the percentage of viable sperm in a sample of the semen of the male patient after the treatment is equal to or higher than a reference value. Preferably, the percentage of viable sperm is equal to or higher than the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0219] 「 Percentage of viable sperm The expression " " is provided above. The method for determining the percentage of viable sperm in a sample is provided above in the definition of asthenozoospermia.

[0220] 「 Percentage of viable sperm in semen samples above the reference value When used herein, the expression " " refers to any percentage of viable sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0221] 「 Percentage of viable sperm in semen samples equal to the reference valueThe expression "」", when used in this specification, refers to any percentage of viable sperm that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than the reference value, where 100% represents the reference value. This also refers to any percentage of viable sperm that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than the reference value, where 100% represents the reference value.

[0222] 「 Percentage of viable sperm in semen samples below the reference value The expression "」", when used in this specification, refers to the percentage of viable sperm in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, preferably at least 25%, more preferably at least 50%, even more preferably at least 100% lower than the reference value, where 100% corresponds to the reference value.

[0223] In certain embodiments, the reference value for the percentage of viable sperm in a sample is 30%, 35%, 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 62%, 64%, 65%, 70%, 75%, preferably 58%. In a preferred embodiment, the reference value for the percentage of viable sperm in a semen sample is the LRL established by the WHO in "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition, which is 58%.

[0224] In another preferred embodiment, the reference value for the percentage of viable sperm in a semen sample from a male subject is the percentage of viable sperm in the semen sample of the male patient obtained prior to the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the percentage of viable sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 samples, preferably at least 2 samples of the semen of the male patient obtained before the treatment referred to in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the percentage of viable sperm in two or more samples obtained before the start of the treatment referred to in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "percentage of viable sperm in the semen sample" obtained in each of the samples. In another embodiment, the samples are each samples obtained at different times before the treatment referred to in the first aspect of the present invention is initiated.

[0225] The terms "post-treatment" and "pre-treatment" are already defined.

[0226] In a particular embodiment of the first aspect of the present invention, the concentration of white blood cells in a semen sample of a male patient after the treatment referred to in the medical use of the present invention is below the reference value. Preferably, the concentration of white blood cells is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples, preferably at least 2 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention.

[0227] In a preferred embodiment, the male patient for the medical use of the present invention is as defined in any of the embodiments of the first aspect of the present invention. Preferably, the patient has pyospermia, and the concentration of white blood cells in the semen sample of the male patient after the treatment referred to in the medical use of the present invention is below the reference value. Preferably, the concentration of white blood cells is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0228] In an embodiment, the male patient is as defined in any of the embodiments of the first aspect of the present invention before the treatment referred to in the above aspect of the present invention. Preferably, the patient has pyospermia before the treatment, and the concentration of white blood cells in the semen sample of the male patient after the treatment is below the reference value. Preferably, the concentration of white blood cells is below the reference value in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20 samples of the semen of the male patient after the treatment referred to in the medical use of the present invention, preferably in at least 2 samples.

[0229] The method for determining the concentration of white blood cells in a semen sample is provided above in the definition of pyospermia.

[0230] “ In semen samples above the reference valueThe expression "concentration of white blood cells", as used herein, refers to any concentration of white blood cells in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than the reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 100%, and still more preferably at least 150% higher, where 100% corresponds to the reference value.

[0231] 「 In semen samples equal to the reference value The expression "concentration of white blood cells", as used herein, refers to any concentration of white blood cells in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% higher than its reference value, preferably less than 5%, more preferably less than 2%, and still more preferably less than 1% higher, where 100% represents the reference value. This also refers to any concentration of white blood cells in a semen sample that is less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, less than 0.05%, less than 0.01%, less than 0.001% lower than its reference value, preferably less than 5%, more preferably less than 2%, and still more preferably less than 1% lower, where 100% represents the reference value.

[0232] 「 In semen samples below the reference value The expression "concentration of white blood cells", as used herein, refers to the concentration of white blood cells in a semen sample that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than its reference value, preferably at least 25%, more preferably at least 50%, even more preferably at least 90% lower, where 100% corresponds to the reference value.

[0233] In certain embodiments, the reference value for the concentration of white blood cells in a semen sample is 1×10 5 , 2.5×10 5 , 5×10 5 , 7.5×10 5 , 1×10 6 , 1.25×10 6 , 1.5×10 6 , 1.75×10 6 , 2×10 6 , 1.25×10 6 , 2.5×10 6 , 2.75×10 6 , 3×10 6 , 5×10 6 white blood cells per milliliter of semen, preferably, 1×10 6 white blood cells per milliliter of semen. In a preferred embodiment, the reference value for the concentration of white blood cells in a semen sample corresponds to the LRL established by the WHO in "Human Semen Examination and Procedures WHO Laboratory Manual", 2010, 5th edition, for the total concentration of white blood cells per milliliter of semen, which is 1×10 6 white blood cells per milliliter of semen, or 1×10 6 peroxidase-positive white blood cells per milliliter of semen.

[0234] In another preferred embodiment, the reference value for the concentration of white blood cells in a semen sample of a male subject is the concentration of white blood cells in the semen sample of the male patient obtained before the treatment referred to in the first aspect of the present invention. In another preferred embodiment, this refers to the concentration of white blood cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, more preferably at least 2 samples of the semen of the male patient obtained before the treatment referred to in the first aspect of the present invention is initiated. In one embodiment, when the reference value refers to the concentration of white blood cells in two or more samples obtained before the start of the treatment referred to in the first aspect of the present invention, the reference value corresponds to the average of the values of the parameter "concentration of white blood cells" obtained in each of the samples. In one embodiment, the samples are each samples obtained at different times before the treatment referred to in the first aspect of the present invention is initiated.

[0235] In a particular embodiment, the type of white blood cells analyzed in the semen sample is polymorphonuclear leukocytes. In this case, the parameter analyzed is the concentration of polymorphonuclear leukocytes instead of the concentration of white blood cells in the semen sample. In this case, all of the above-provided embodiments in which the expression "concentration of white blood cells in the semen sample" is used are also applied by replacing the expression "concentration of white blood cells" with "concentration of polymorphonuclear leukocytes". The reference value for the polymorphonuclear leukocyte concentration is 0.5×10 5 、1×10 5 、2.5×10 5 、5×10 5 、7.5×10 5 、1×10 6 、1.25×10 6 、1.5×10 6 、1.75×10 6 、2×10 6 、1.25×10 6 、2.5×10 6 、2.75×10 6 、3×10 6 、5×10 6Polymorphonuclear leukocytes per milliliter of semen, preferably, 5×10 5 Polymorphonuclear leukocytes per milliliter of semen.

[0236] The terms "post-treatment" and "pre-treatment" are already defined.

[0237] In a preferred embodiment, the treatment referred to in the first aspect of the present invention comprises administration of the biomass, protein extract or pharmaceutical composition of the first aspect of the present invention at a certain period, a certain frequency, and a certain dose at each time during this period.

[0238] In certain embodiments, the period is at least 7 days, at least 15 days, at least 20 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 50 days, at least 60 days, at least 65 days, at least 70 days, at least 75 days, at least 80 days, at least 85 days, at least 90 days, at least 95 days, at least 100 days, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 1.5 years, at least 2 years, at least 3 years, at least 5 years, at least 10 years, preferably at least 30 days (i.e., 1 month), more preferably at least 65 days, even more preferably at least 70 days, and still more preferably at least 90 days. Thus, in certain embodiments, the treatment referred to in the medical use of the present invention comprises administration of the biomass, protein extract or pharmaceutical composition referred to in the first aspect of the present invention for at least 30 days (i.e., 1 month). In another particular embodiment, the treatment referred to in the medical use of the present invention comprises administration of the biomass, protein extract or pharmaceutical composition referred to in the first aspect of the present invention for at least 65 days. In another particular embodiment, the treatment referred to in the medical use of the present invention comprises administration of the biomass, protein extract or pharmaceutical composition referred to in the first aspect of the present invention for at least 70 days. In another particular embodiment, the treatment referred to in the medical use of the present invention comprises administration of the biomass, protein extract or pharmaceutical composition referred to in the first aspect of the present invention for at least 90 days (i.e., 3 months).

[0239] In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day every day during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every two days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every three days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every four days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every five days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every six days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every seven days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every ten days during the period. In another specific embodiment, the frequency is preferably at least once a day, at least twice a day, at least three times a day, at least four times a day, at least five times a day once every two weeks during the period. In a preferred embodiment, the frequency is once a day during the period. In a more preferred embodiment, the frequency is once a day, preferably every day, during the entire period of treatment at any of the periods shown above.

[0240] In certain embodiments, when the treatment referred to in the first aspect of the present invention includes the administration of the biomass of T. chuii, the biomass is dehydrated biomass, and the dose at which the biomass is administered is 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1 g, 1.5 g, 2 g, 2.5 g, 3 g, 3.5 g, 4 g, 4.5 g, 5 g of dehydrated biomass. In another preferred embodiment, the treatment referred to in the first aspect of the present invention includes administering 10 to 500 mg of dehydrated biomass, more preferably 500 mg, and even more preferably 250 mg of dehydrated biomass of T. chuii. In a preferred embodiment, the treatment of the first aspect of the present invention includes administering 10 to 500 mg of dehydrated biomass per day, preferably 250 mg of dehydrated biomass per day.

[0241] In another specific embodiment, when the biomass is fresh biomass, the dosage is 2.5 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 290 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 625 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1 g, 1.25 g, 1.5 g, 1.7 g, 2 g, 2.1 g, 2.2 g, 2.3 g, 2.4 g, 2.5 g, 2.6 g, 2.7 g, 2.8 g, 2.9 g, 3 g, 3.25 g, 3.5 g, 3.75 g, 4 g, 4.5 g, 5 g of fresh biomass.

[0242] In a preferred embodiment, the treatment referred to in the first aspect of the present invention comprises administering 30 mg to 4 g, preferably 4 g, more preferably 3 g, even more preferably 40 mg, and still more preferably 30 mg of fresh biomass. In another preferred embodiment, the treatment referred to in the first aspect of the present invention comprises administering 30 mg to 4 g, preferably 4 g, more preferably 3 g, even more preferably 40 mg, and still more preferably 30 mg of fresh biomass per day.

[0243] In certain embodiments, the treatment referred to in the first aspect of the present invention comprises administration of a protein extract of T. thuisii as defined herein, and the dosage for such administration is 0.2 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.8 mg, 1 mg, 1.2 mg, 1.4 mg, 1.6 mg, 1.8 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 8 mg, 10 mg, 12 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1 g, 1.5 g, 2 g of the protein extract of T. thuisii.

[0244] In another preferred embodiment, the treatment referred to in the first aspect of the present invention comprises administering 4 - 200 mg of the protein extract, more preferably 200 mg, and even more preferably 100 mg of the protein extract of T. thuisii. In a preferred embodiment, the treatment of the first aspect of the present invention comprises administering 4 - 200 mg of the protein extract per day, preferably 10 mg of the protein extract per day.

[0245] In certain embodiments, when the treatment referred to in the first aspect of the present invention comprises administration of a pharmaceutical composition comprising fresh biomass of T. thuisii, this is administered in a dosage that results in administration of the fresh biomass in any of the amounts shown above for the fresh biomass. In another particular embodiment, when the pharmaceutical composition comprises dehydrated biomass of T. thuisii, this is administered in a dosage that results in administration of the dehydrated biomass in any of the amounts shown above for the dehydrated biomass. In another particular embodiment, when the treatment referred to in the first aspect of the present invention comprises administration of a pharmaceutical composition comprising a protein extract of T. thuisii, this is administered in a dosage that results in administration of the protein extract in any of the amounts shown above for the protein extract of T. thuisii.

[0246] In a preferred embodiment, the biomass, protein or pharmaceutical composition referred to in the medical use of the present invention is in the form of tablets, capsules, liquid suspensions, dry powders, or wet compositions. Thus, in a preferred embodiment, the biomass, protein extract or pharmaceutical composition referred to in the medical use of the present invention is administered in any of solid forms.

[0247] II - Method of the present invention In a second aspect, the present invention relates to a method for improving the semen quality of a male subject who has normal semen, does not have oligospermia, and does not have a disorder characterized by a percentage of sperm DNA fragmentation (SDF) above a reference value, the method comprising the step of treating the subject by administering a biomass of T. tuyili or a protein extract of T. tuyili. In certain embodiments, the male subject of the second aspect of the present invention does not have a disorder characterized by a level of 8-OH-dG in semen above a reference value, an 8OH-dG / 10 5 dG ratio, or a percentage of 8-OH-dG positive sperm cells. In another particular embodiment, the male subject of the second aspect of the present invention does not have a disorder characterized by sORP in semen above a reference value. In certain embodiments, the male subject of the second aspect of the present invention does not have azoospermia. In another particular embodiment, the male subject of the second aspect of the present invention does not have pyospermia. In another particular embodiment, the male subject of the second aspect of the present invention does not have necrospermia. In another particular embodiment, the male subject of the second aspect of the present invention has normal semen, does not have oligospermia, does not have a disorder characterized by a percentage of SDF above a reference value, does not have a disorder characterized by a level of 8-OH-dG in semen above a reference value, an 8OH-dG / 10 5Does not have a disorder characterized by the ratio of dG or the percentage of 8-OH-dG positive sperm cells, does not have a disorder characterized by sORP in semen above the reference value, does not have azoospermia, does not have pyospermia, and does not have necrospermia. In certain embodiments, the male subject of the second aspect of the invention does not suffer from infertility.

[0248] In certain embodiments, the method of the second aspect is a non-therapeutic method.

[0249] 「 Semen quality of male subjects The expression " " as used herein refers to the characterization of the semen of a male subject with respect to a method that achieves fertilization excellently or well. The quality of semen is determined by measuring different semen parameters. Non-limiting examples of said parameters include the following: total number of sperm in a semen sample, concentration of sperm in a semen sample, volume of semen obtained from ejaculated semen, percentage of morphologically normal sperm in a semen sample, percentage of progressively motile sperm in a semen sample, percentage of viable sperm in a semen sample, concentration of white blood cells in a semen sample, percentage of SDF, level of 8-OH-dG in a semen sample, 8OH-dG / 10 5 ratio of dG or percentage of 8-OH-dG positive sperm cells, or sORP in a semen sample.

[0250] "azoospermia", "oligospermia", "pyospermia", "necrospermia", "morphologically normal sperm", "progressively motile sperm", "percentage of SDF", "level of 8-OH-dG in semen", "8OH-dG / 10 5The definitions of "ratio of dG", "percentage of 8-OH-dG positive sperm cells in semen", and "sORP in semen" are provided in the first aspect of the present invention and applied to the second aspect of the present invention. The total number of sperm in the semen sample, the concentration of sperm in the semen sample, the volume of semen obtained from the ejaculated semen, the percentage of morphologically normal sperm in the semen sample, the percentage of progressively motile sperm in the semen sample, the percentage of viable sperm in the semen sample, the concentration of white blood cells in the semen sample, the percentage of SDF, the level of 8-OH-dG in the semen sample, 8OH-dG / 10 5 A method for determining the ratio of dG or the percentage of 8-OH-dG positive sperm cells, or sORP in a semen sample is also provided in the first aspect of the present invention and applied to the second aspect of the present invention.

[0251] 「 The semen quality of male subjects Improvement Cause 」 or 「 The semen quality of male subjects Improvement Causing 」 refers to an improvement in the ability of the semen of the male subject to achieve fertilization as compared to a control semen sample from the male subject. The improvement can be determined by measuring at least one semen parameter in the semen sample from the male subject, where if at least one semen parameter in the semen sample shows improvement compared to the control semen sample, it is considered that the quality of the semen of the male subject has improved, that is, the ability of the semen of the male subject to achieve fertilization has improved. Therefore, 「 Method for improving the semen quality of male subjects 」, when used herein, refers to a method for improving at least one semen parameter in the semen of a male subject compared to a control sample.

[0252] In a preferred embodiment, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 samples of the male subject, preferably at least 2 semen samples, must show an improvement in at least one semen parameter with respect to the control sample for the quality of the semen of the male subject that is considered to be improved. Thus, in a preferred embodiment, a method for improving the quality of semen of a male subject, as used herein, refers to a method for improving at least one semen parameter in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10 semen samples of the male subject, preferably at least 2 semen samples, with respect to the control sample.

[0253] The semen parameters in a semen sample of a male subject are such that when the semen parameter is the total number of sperm in the semen sample, the volume of semen obtained from the ejaculate, the percentage of morphologically normal sperm in the semen sample, the percentage of progressively motile sperm in the semen sample, or the percentage of viable sperm in the semen sample, and the value of the parameter is higher in the semen sample of the male subject than the value of the parameter in the control semen sample, it is considered improved with respect to the control semen sample. The semen parameters in a semen sample of a male subject are also such that when the parameter is the concentration of white blood cells in the semen sample, the percentage of SDF in the semen sample, the level of 8OH-dG in the semen sample, the ratio of 8OH-dG / 10 5 dG in the semen sample, the percentage of 8-OH-dG positive sperm cells in the semen sample, or the sORP in the semen sample, and the value of the parameter is lower in the semen sample of the male subject than the value of the parameter in the control sample, it is considered improved with respect to the control semen sample.

[0254] In a preferred embodiment, " The value of the parameter is higher in the semen sample of the male subject than the value of the parameter in the control semen sampleAs used herein, the expression "」 refers to the value of the parameter in the semen sample of a male subject that is at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, at least 150%, at least 200%, at least 300%, at least 400%, at least 500%, at least 600%, at least 800%, or at least 1000% higher than in the control semen sample. In a preferred embodiment, the expression refers to the value of the parameter in the semen sample of a male subject that is at least 25% higher than the value of the parameter in the control sample. In another preferred embodiment, the expression refers to the value of the parameter in the semen sample of a male subject that is at least 50% higher than the value of the parameter in the control sample. In another preferred embodiment, the expression refers to the value of the parameter in the semen sample of a male subject that is at least 100% higher than the value of the parameter in the control sample. In another preferred embodiment, the expression refers to the value of the parameter in the semen sample of a male subject that is at least 150% higher than the value of the parameter in the control sample. In another preferred embodiment, the expression refers to the value of the parameter in the semen sample of a male subject that is at least 200% higher than the value of the parameter in the control sample.

[0255] In another preferred embodiment, " The value of the parameter is lower in the semen sample of the male subject than the value of the parameter in the control sampleAs used herein, the expression "」" refers to the value of the parameter in a semen sample of a male subject that is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 90%, at least 100% lower than that in a control sample. In a preferred embodiment, the expression refers to the value of the parameter in a semen sample of a male subject that is at least 25% lower than the value of the parameter in the control sample. In a preferred embodiment, the expression refers to the value of the parameter in a semen sample of a male subject that is at least 50% lower than the value of the parameter in the control sample. In a preferred embodiment, the expression refers to the value of the parameter in a semen sample of a male subject that is at least 80% lower than the value of the parameter in the control sample. In a preferred embodiment, the expression refers to the value of the parameter in a semen sample of a male subject that is at least 90% lower than the value of the parameter in the control sample. In a preferred embodiment, the expression refers to the value of the parameter in a semen sample of a male subject that is at least 100% lower than the value of the parameter in the control sample.

[0256] In one embodiment, when the control sample is a set of samples shown below, the value of the semen parameter in the control sample is the average of the values of the parameter in each of the samples in the set.

[0257] In a preferred embodiment, the control sample is a semen sample from a male subject of the method of the present invention obtained at least 1 hour before, at least 2 hours before, at least 3 hours before, at least 4 hours before, at least 5 hours before, at least 6 hours before, at least 7 hours before, at least 8 hours before, at least 9 hours before, at least 10 hours before, at least 11 hours before, at least 12 hours before, at least 18 hours before, at least 1 day before, at least 2 days before, at least 3 days before, at least 5 days before, at least 6 days before, at least 1 week before, at least 2 weeks before, at least 3 weeks before, at least 1 month before, at least 2 months before, at least 3 months before, at least 6 months before, at least 9 months before, at least 1 year before, at least 2 years before, at least 3 years before, at least 4 years before, at least 5 years before the method of the present invention is performed. In a more preferred embodiment, the control sample is a semen sample from a male subject of the method of the present invention obtained preferably at least 1 month before, more preferably at least 1 week before, even more preferably at least 1 day before, and still more preferably at least 1 hour before the method of the present invention is performed.

[0258] In one embodiment, the control sample is a set of semen samples from a male subject of the method of the present invention, preferably a set of 2, 3, 4, or 5, more preferably a set of 2 semen samples. In a preferred embodiment, they are obtained at different time points selected from those shown above for the control sample. In this case, the sample is obtained as described in any of the embodiments provided in the first aspect of the present invention for obtaining a semen sample.

[0259] In a preferred embodiment, the control sample is from a subject having normal semen. In another preferred embodiment, the control sample is from a subject not having a disorder associated with a percentage of SDF higher than the reference value, i.e., the control sample exhibits a percentage of SDF lower than the reference value. In another specific embodiment, the control sample is from a subject not having oligospermia. In a specific embodiment, when the control sample is the set of semen samples shown above, each of the samples has normal semen, does not have oligospermia, and / or is from a male subject not having a disorder associated with a percentage of SDF higher than the reference value, i.e., the control sample exhibits a percentage of SDF lower than the reference value.

[0260] In another embodiment, the control sample is from a subject not having a disorder associated with the level of 8-OH-dG, the ratio of 8OH-dG / 10 5 dG, or the percentage of 8-OH-dG positive sperm cells in semen above the reference value, i.e., the control sample exhibits a level of 8-OH-dG, a ratio of 8OH-dG / 10 5 dG, or a percentage of 8-OH-dG positive sperm cells lower than the reference value. In another specific embodiment, the control sample is from a subject not having a disorder associated with sORP in semen above the reference value, i.e., the control sample exhibits an sORP lower than the reference value. In another specific embodiment, the control sample is from a subject not having necrospermia. In another specific embodiment, the control sample is from a subject not having pyospermia. As will be understood by those skilled in the art, the control sample is from a subject not having azoospermia.

[0261] “ Normal semen ” As used herein, refers to a state characterized in that the total number of sperm and / or the concentration of sperm, the percentage of PR, and the percentage of morphologically normal sperm in the semen of a male subject are above the corresponding LRL. The WHO has established the LRR for the following in the "WHO Laboratory Manual for the Examination and Processing of Human Semen", 2010, 5th Edition: - The total sperm count is 39×10 per ejaculate, 6 and, - The sperm concentration is 15×10 per ml, 6 and, - The percentage of progressively motile sperm is 32%, - The percentage of morphologically normal sperm is 4%.

[0262] In a preferred embodiment, the subject has normal semen when at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject show a total sperm count and / or sperm concentration, percentage of PR, and percentage of morphologically normal sperm that are above the corresponding reference values. In another preferred embodiment, the reference values for the total sperm count and / or sperm concentration, percentage of PR, and percentage of morphologically normal sperm are any of those provided for each of the parameters in the first aspect of the present invention.

[0263] A method for determining the total sperm count, sperm concentration, percentage of PR, and percentage of morphologically normal sperm in the semen of a male patient or in a semen sample is provided in the first aspect of the present invention in the definitions of "oligospermia", "asthenospermia", and "teratospermia" and is applicable to the second aspect of the present invention.

[0264] In a preferred embodiment, the reference value for the percentage of SDF referred to in the second aspect of the present invention is any of the reference values provided for the parameters in the first aspect of the present invention. A method for determining the percentage of SDF in the semen of a male patient or in a semen sample is provided in the first aspect of the present invention in the definition of "percentage of SDF".

[0265] In another preferred embodiment, the reference value for the level of 8-OH-dG in semen or in a semen sample, the ratio of 8OH-dG / 10 5 dG or the percentage of 8-OH-dG positive sperm cells, as referred to in the second aspect of the present invention, is any of the reference values provided for said parameters in the first aspect of the present invention. The method for determining the level of 8-OH-dG in semen or in a semen sample, the ratio of 8OH-dG / 10 5 dG or the percentage of 8-OH-dG positive sperm cells in a male patient is provided in the first aspect of the present invention, respectively, in the definitions of "level of 8-OH-dG in semen", "8OH-dG / 10 5 ratio of dG" or "percentage of 8-OH-dG positive sperm cells in semen".

[0266] In a preferred embodiment, the reference value of sORP as referred to in the second aspect of the present invention is any of the reference values provided for said parameters in the first aspect of the present invention. The method for determining sORP in semen or in a semen sample of a male patient is provided in the first aspect of the present invention according to the definition of "sORP in semen".

[0267] In another preferred embodiment, the male subject of the second aspect of the present invention is fertile.

[0268] " Fertility ", when used herein, refers to the genital state defined by the ability to achieve a clinical pregnancy within 12 months or less of regular unprotected sexual intercourse. " Male fertilityThe expression "」", when used in this specification, refers to the reproductive ability of male subjects. As understood by those skilled in the art, a male showing reproductive ability is preferably characterized by not showing changes leading to infertility, particularly deficiencies in any semen parameters. Said semen parameters include the total number of sperm, sperm concentration, volume of semen obtained from ejaculate, percentage of morphologically normal sperm, percentage of progressively motile sperm, percentage of viable sperm, concentration of white blood cells, percentage of SDF, level of 8-OH-dG, ratio of 8OH-dG / 10 5 dG or percentage of 8-OH-dG positive sperm cells, or level of sORP. As understood by those skilled in the art, a semen sample of a male subject considered to have reproductive ability preferably shows a number and / or concentration of sperm, volume of semen per ejaculate, percentage of morphologically normal sperm, percentage of progressively motile sperm and percentage of viable sperm that are above the reference values. In addition, a semen sample of a male subject considered to have reproductive ability preferably shows a concentration of white blood cells, percentage of SDF, level of 8-OH-dG, 8OH-dG / 10 5 dG ratio, percentage of 8-OH-dG positive sperm cells and level of sORP that are below the reference values. Thus, in certain embodiments, a semen sample of a male subject of the method of the present invention shows a total number of sperm, sperm concentration, volume of semen per ejaculate, percentage of morphologically normal sperm, percentage of progressively motile sperm and percentage of viable sperm that are above the reference values. In addition, a semen sample of a male subject considered to have reproductive ability preferably shows a concentration of white blood cells, percentage of SDF, level of 8-OH-dG, 8OH-dG / 10 5 dG ratio, percentage of 8-OH-dG positive sperm cells and level of sORP that are below the reference values.

[0269] In a preferred embodiment, when the male subject is fertile, at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the subject's semen have a total sperm count, sperm concentration, semen volume per ejaculate, percentage of morphologically normal sperm, percentage of progressively motile sperm and percentage of viable sperm that are above a reference value, and a white blood cell concentration, percentage of SDF, level of 8-OH-dG, ratio of 8OH-dG / 10 5 dG ratio, percentage of 8-OH-dG positive sperm cells and level of sORP that are below the reference value.

[0270] The expressions "total sperm count in a semen sample below the reference value", "sperm concentration in a semen sample below the reference value", "percentage of morphologically normal sperm in a semen sample below the reference value", "percentage of progressively motile sperm in a semen sample below the reference value", "semen volume per ejaculate in a semen sample below the reference value", "percentage of SDF in a semen sample above the reference value", "level of 8-OH-dG in a semen sample above the reference value", "ratio of 8OH-dG / 10 5 dG ratio in a semen sample above the reference value", "percentage of 8-OH-dG positive sperm cells in a semen sample above the reference value", "sORP above the reference value", "white blood cell concentration in a semen sample above the reference value", and "percentage of viable sperm below the reference value in a semen sample" are defined and described in the different embodiments above in the first aspect of the invention, and the definitions and embodiments apply to the second aspect of the invention.

[0271] "Total number of spermatozoa in a semen sample equal to the reference value", "Sperm concentration in a semen sample equal to the reference value", "Percentage of morphologically normal spermatozoa in a semen sample equal to the reference value", "Percentage of progressively motile spermatozoa in a semen sample equal to the reference value", "Volume of semen per ejaculate in a semen sample equal to the reference value", "Percentage of SDF in a semen sample equal to the reference value", "Level of 8-OH-dG in a semen sample equal to the reference value", "Ratio of 8OH-dG / 10 5 dG in a semen sample equal to the reference value", "Percentage of 8-OH-dG positive sperm cells in a semen sample equal to the reference value", "sORP equal to the reference value", "Concentration of white blood cells in a semen sample equal to the reference value", and "Percentage of viable spermatozoa in a semen sample equal to the reference value" are defined and described in the above different embodiments in the first aspect of the present invention, and the said definitions and embodiments are applicable to the second aspect of the present invention.

[0272] "Total number of spermatozoa in a semen sample exceeding the reference value", "Sperm concentration in a semen sample exceeding the reference value", "Percentage of morphologically normal spermatozoa in a semen sample exceeding the reference value", "Percentage of progressively motile spermatozoa in a semen sample exceeding the reference value", "Volume of semen per ejaculate in a semen sample exceeding the reference value", "Percentage of SDF in a semen sample below the reference value", "Level of 8-OH-dG in a semen sample below the reference value", "Ratio of 8OH-dG / 105dG in a semen sample below the reference value", "Percentage of 8-OH-dG positive sperm cells in a semen sample below the reference value", "sORP below the reference value", "Concentration of white blood cells in a semen sample below the reference value", and "Percentage of viable spermatozoa in a semen sample exceeding the reference value" are defined and described in the above different embodiments in the first aspect of the present invention, and the said definitions and embodiments are applicable to the second aspect of the present invention.

[0273] In a preferred embodiment, the reference values for the total number of sperm in the semen sample, the concentration of sperm in the semen sample, the percentage of morphologically normal sperm in the semen sample, the percentage of progressively motile sperm in the semen sample, the volume of semen per ejaculate in the semen sample, the percentage of SDF in the semen sample, the level of 8-OH-dG in the semen sample, the ratio of 8OH-dG / 10 5 dG in the semen sample, the percentage of 8-OH-dG positive sperm cells in the semen sample, the level of sORP in the semen sample, the concentration of white blood cells in the semen sample, and the percentage of viable sperm in the semen sample are those specified in the first aspect of the present invention for the corresponding parameters.

[0274] The term "treating" as used in the second aspect of the present invention refers to the performance of a treatment as defined in the first aspect of the present invention and described in any of the embodiments of the first aspect of the present invention, where the treatment is non-therapeutic. Thus, as will be understood by those skilled in the art, the treatment of the second aspect of the present invention applied to a male subject is as described in the first aspect of the present invention, where there is no disease or disorder such as infertility or any semen disorder that is being treated or to be treated in the male subject by said treatment.

[0275] In one embodiment, the number and / or concentration of sperm in the semen sample of the male subject of the method of the present invention obtained after the treatment referred to in the method is higher than that in the semen sample of the male subject obtained before the treatment.

[0276] In a preferred embodiment, the number and / or concentration of sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment referred to in the method is higher than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0277] In one embodiment, the percentage of progressively motile sperm in a semen sample of a male subject of the method of the present invention obtained after the treatment referred to in the method is higher than that in a sample of the semen of the male subject obtained before the treatment.

[0278] In a preferred embodiment, the percentage of progressively motile sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment referred to in the method is higher than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0279] In one embodiment, the percentage of morphologically normal sperm in a semen sample of a male subject of the method of the present invention obtained after the treatment referred to in the method is higher than that in a sample of the semen of the male subject obtained before the treatment.

[0280] In a preferred embodiment, the percentage of morphologically normal sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment mentioned in the method is higher than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0281] In one embodiment, the volume of semen per ejaculate in a semen sample of a male subject of the method of the present invention obtained after the treatment mentioned in the method is higher than that in a sample of the semen of the male subject obtained before the treatment.

[0282] In a preferred embodiment, the volume of semen per ejaculate in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment mentioned in the method is higher than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0283] In one embodiment, the percentage of SDF in a sample of semen of a male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in a sample of the semen of the male subject obtained before the treatment.

[0284] In a preferred embodiment, the percentage of SDF in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment referred to in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0285] In one embodiment, the level of 8-OH-dG in a sample of semen of a male subject of the method of the present invention obtained after the treatment referred to in the method is lower than that in a sample of semen of the male subject obtained before the treatment.

[0286] In a preferred embodiment, the level of 8-OH-dG in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment referred to in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0287] In one embodiment, 8-OH-dG / 10 5 dG ratio in a sample of semen of a male subject of the method of the present invention obtained after the treatment referred to in the method is lower than that in a sample of semen of the male subject obtained before the treatment.

[0288] In a preferred embodiment, the 8-OH-dG / 10 5 dG ratio in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0289] In one embodiment, the percentage of 8-OH-dG positive sperm cells in a sample of semen of a male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in a sample of semen of the male subject obtained before the treatment.

[0290] In a preferred embodiment, the percentage of 8-OH-dG positive sperm cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of a male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0291] In one embodiment, the sORP in a sample of semen of a male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in a sample of semen of the male subject obtained before the treatment.

[0292] In a preferred embodiment, the sORP in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0293] In one embodiment, the concentration of white blood cells in the semen sample of the male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in the sample of the semen of the male subject obtained before the treatment.

[0294] In a preferred embodiment, the concentration of white blood cells in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject of the method of the present invention obtained after the treatment mentioned in the method is lower than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0295] In one embodiment, the percentage of viable sperm in the semen sample of the male subject of the method of the present invention obtained after the treatment mentioned in the method is higher than that in the sample of the semen of the male subject obtained before the treatment.

[0296] In a preferred embodiment, the percentage of viable sperm in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 semen samples of the male subject of the method of the present invention obtained after the treatment mentioned in the method is higher than that in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, preferably at least 2 samples of the semen of the male subject obtained before the treatment.

[0297] In one embodiment, when referring to the parameters of "sperm count", "sperm concentration", "percentage of progressively motile sperm", "percentage of morphologically normal sperm", "volume of semen per ejaculate", "percentage of viable sperm" in a semen sample obtained after the treatment mentioned in the method of the present invention Higher than that in the semen sample of the male subject obtained before the treatment The expression of "

[0298] In one embodiment, when referring to the parameters of "percentage of SDF", "level of 8-OH-dG", "8-OH-dG / 10 5 dG ratio", "percentage of 8-OH-dG positive sperm cells", "sORP" and "concentration of white blood cells" in a semen sample obtained after the treatment mentioned in the method of the present invention Lower than that in the semen sample of the male subject obtained before the treatmentThe expression "..." is understood to mean that the value of the corresponding parameter in at least the semen sample is at least 5%, at least 10%, at least 25%, at least 40%, at least 50%, at least 80%, at least 100%, preferably at least 25%, more preferably at least 50%, even more preferably at least 100% lower than the value of the parameter in the semen sample(s) obtained before the said treatment.

[0299] In certain embodiments, the sample obtained after the treatment referred to in the method of the present invention is obtained at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 18 hours, at least 1 day, at least 2 days, at least 3 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, at least 5 years after the said treatment. In a more preferred embodiment, the sample obtained after the treatment referred to in the method of the present invention is obtained at least 1 month, more preferably at least 1 week, even more preferably at least 1 day, and still more preferably at least 1 hour after the said treatment.

[0300] In one embodiment, the plurality of samples obtained after the treatment referred to in the method of the present invention are obtained at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 18 hours, at least 1 day, at least 2 days, at least 3 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, at least 5 years after the treatment. In a more preferred embodiment, the plurality of samples obtained after the treatment referred to in the method of the present invention are obtained at least 1 month, more preferably at least 1 week, even more preferably at least 1 day, and still more preferably at least 1 hour after the treatment.

[0301] In one embodiment, the sample obtained before the treatment referred to in the method of the present invention is obtained at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 18 hours, at least 1 day, at least 2 days, at least 3 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 9 months, at least 1 year, at least 2 years, at least 3 years, at least 4 years, at least 5 years before the treatment. In a more preferred embodiment, the sample obtained before the treatment referred to in the method of the present invention is obtained at least 1 month, more preferably at least 1 week, even more preferably at least 1 day, and still more preferably at least 1 hour before the treatment.

[0302] In one embodiment, the plurality of samples obtained prior to the treatment referred to in the method of the present invention are obtained at least 1 hour before, at least 2 hours before, at least 3 hours before, at least 4 hours before, at least 5 hours before, at least 6 hours before, at least 7 hours before, at least 8 hours before, at least 9 hours before, at least 10 hours before, at least 11 hours before, at least 12 hours before, at least 18 hours before, at least 1 day before, at least 2 days before, at least 3 days before, at least 5 days before, at least 6 days before, at least 1 week before, at least 2 weeks before, at least 3 weeks before, at least 1 month before, at least 2 months before, at least 3 months before, at least 6 months before, at least 9 months before, at least 1 year before, at least 2 years before, at least 3 years before, at least 4 years before, at least 5 years before the said treatment. In a more preferred embodiment, the plurality of samples obtained prior to the treatment referred to in the method of the present invention are obtained at least 1 month before, more preferably at least 1 week before, even more preferably at least 1 day before, and still more preferably at least 1 hour before the said treatment.

[0303] In a particular embodiment, the biomass of T. tuyi referred to in the method of the present invention is dehydrated. In another particular embodiment, the biomass of T. tuyi referred to in the method of the present invention is fresh.

[0304] 「 Fresh biomass of T. thuyii 」and「 Dehydrated biomass of T. thuyii 」are defined in the first aspect of the present invention and are applicable to the second aspect of the present invention.

[0305] As described above, the treatment referred to in the method of the present invention is as per the treatment of the first aspect of the present invention, which is non-therapeutic, and the embodiments provided in the first aspect of the present invention in combination with the treatment of the first aspect of the present invention are the treatment of the second aspect of the present invention, provided that the treatment is applied to a non-therapeutic treatment. Thus, in certain embodiments, the treatment referred to in the method of the present invention includes administering dehydrated biomass at 10 to 500 mg per day, preferably 250 mg per day. In another specific embodiment, this includes administering fresh biomass at 30 mg to 4 g per day, preferably 4 g, more preferably 3 g, even more preferably 40 mg, and still more preferably 30 mg of fresh biomass per day. In another specific embodiment, this includes administering protein extract at 4 to 200 mg per day, preferably 100 mg of protein extract of T. thuisii per day.

[0306] In another preferred embodiment, the treatment referred to in the second aspect of the present invention includes administering at least 30 days (i.e., one month) of the biomass or protein extract as defined herein. In another preferred embodiment, the treatment referred to in the second aspect of the present invention includes administering at least 65 days of the biomass or protein extract as defined herein. In another preferred embodiment, the treatment referred to in the second aspect of the present invention includes administering at least 70 days of the biomass or protein extract as defined herein. In another preferred embodiment, the treatment referred to in the second aspect of the present invention includes administering at least 90 days of the biomass or protein extract as defined herein.

[0307] All terms and embodiments described in the first aspect of the present invention are equally applicable to this aspect of the present invention.

[0308] III - Use of the present invention In a third aspect, the present invention relates to the use of the biomass of T. tuyili or the protein extract of T. tuyili as an oral supplement for improving the quality of semen of male subjects who have normal semen, do not have oligospermia, and do not have a disorder characterized by the percentage of SDF in a semen sample above a reference value. In another specific embodiment, the subject of the third aspect of the present invention does not have a disorder characterized by the level of 8-OH-dG, 8-OH-dG / 10 5 dG ratio, or percentage of 8-OH-dG positive sperm cells in semen above a reference value. In another specific embodiment, the subject of the third aspect of the present invention does not have a disorder characterized by sORP in semen above a reference value. In a specific embodiment, the subject of the third aspect of the present invention does not have azoospermia. In a specific embodiment, the subject of the third aspect of the present invention does not have pyospermia. In another specific embodiment, the subject of the third aspect of the present invention does not have necrospermia. In another specific embodiment, the subject of the third aspect of the present invention has normal semen, does not have oligospermia, does not have a disorder characterized by the percentage of SDF in a semen sample above a reference value, does not have a disorder characterized by the level of 8-OH-dG, 8-OH-dG / 10 5 dG ratio, or percentage of 8-OH-dG positive sperm cells in semen above a reference value, does not have a disorder characterized by the level of sORP in semen above a reference value, does not have azoospermia, does not have pyospermia, and does not have necrospermia.

[0309] 「 Oral supplementAs used herein, the expression "」 refers to any product that is provided with the target meal and administered orally. Oral supplements can be incorporated in a very different form, including tablets, capsules, liquid suspensions, dry powders, wet compositions, enteral nutrition dry or enteral nutrition wet. In a preferred embodiment, the oral supplement comprises or consists of the biomass of T. chuii, the protein extract of T. chuii or the pharmaceutical composition as defined and described in any embodiment of the first aspect of the present invention. In another preferred embodiment, the oral supplement, the biomass of T. chuii or the protein extract of T. chuii referred to in the use of the present invention is administered as shown in any embodiment of the first aspect of the present invention for the administration of the biomass of T. chuii, the protein extract of T. chuii or the pharmaceutical composition that is constituted within the treatment referred to in the first aspect of the present invention. Thus, in certain embodiments, the oral supplement, the biomass of T. chuii or the protein extract of T. chuii referred to in the use of the present invention is administered for the period, frequency and dosage shown in any embodiment of the first aspect of the present invention that specifies the period, frequency and dosage of administration of the biomass, protein extract or pharmaceutical composition referred to in the first aspect of the present invention.

[0310] 「 Semen quality of male subjects 」 and 「 Improve the semen quality of male subjects 」 are as defined and described in any embodiment of the second aspect of the present invention.

[0311] The definitions of the terms "normal semen", "oligospermia", "percentage of SDF", "azoospermia", "pyospermia", "necrospermia", "level of 8-OH-dG in semen", "8-OH-dG / 10 5 dG ratio", "percentage of 8-OH-dG positive sperm cells" and "level of sORP in semen" are also provided in the first or second aspect of the present invention and apply to this aspect of the present invention.

[0312] The expression "percentage of SDF above the reference value" is as defined and described in any embodiment of the first and second aspects of the present invention.

[0313] In a preferred embodiment, the reference value of the percentage of SDF is any of those shown in the first aspect of the present invention.

[0314] The expression "level of 8-OH-dG in semen above the reference value" is as defined and described in any embodiment of the first and second aspects of the present invention.

[0315] In a preferred embodiment, the reference value of the level of 8-OH-dG in semen is any of those shown in the first aspect of the present invention.

[0316] 「The expression "8-OH-dG / 10 5 dG ratio in semen above the reference value" is as defined and described in any embodiment of the first and second aspects of the present invention.

[0317] In a preferred embodiment, the reference value of the 8-OH-dG / 10 5 dG ratio in semen is any of those shown in the first aspect of the present invention.

[0318] The expression "percentage of 8-OH-dG positive sperm cells in semen above the reference value" is as defined and described in any embodiment of the first and second aspects of the present invention.

[0319] In a preferred embodiment, the reference value of the percentage of 8-OH-dG positive sperm cells in semen is any of those shown in the first aspect of the present invention.

[0320] The expression "sORP in semen above the reference value" is as defined and described in any embodiment of the first and second aspects of the present invention.

[0321] In a preferred embodiment, the reference value of sORP in semen is any of those shown in the first aspect of the present invention.

[0322] In a specific embodiment, the male subject of the third aspect of the present invention is the same as the male subject of the second aspect of the present invention. In a preferred embodiment, the male subject of the third aspect of the present invention is fertile.

[0323] The expression "fertile" is as defined and described in any of the embodiments of the second aspect of the present invention.

[0324] In one embodiment, all terms and embodiments described in the first and second aspects of the present invention are equally applicable to this aspect of the present invention.

[0325] In one embodiment, the terms "comprising" and "consisting of" used in the first, second and third aspects of the present invention are interchangeable.

[0326] The present invention will be described by the following examples, which are merely illustrative of the present invention and are not to be considered as limiting the scope of the present invention.

Examples

[0327] Materials and methods A total of 20 individuals showing idiopathic oligospermia, asthenozoospermia and / or teratozoospermia (by the first complete semen analysis) were recruited. Each of these conditions was defined according to the World Health Organization (WHO) parameters described in the "WHO Laboratory Manual for the Examination and Processing of Human Semen" (5th Edition, 2010). More specifically, the term oligospermia was defined as 39×10 6 sperm per ejaculate (or 15×10 6Refers to the total number of sperm (or concentration depending on the reported result) below the lower reference limit (LRL) of sperm / ml. Regarding asthenozoospermia, this occurs when the percentage of progressively motile (PR) sperm is below the LRL (32%). Finally, teratozoospermia is diagnosed when the percentage of morphologically normal sperm is below the LRL (4%). Regarding this nomenclature, PR refers to sperm that are actively moving either linearly or in large circles regardless of speed, and for sperm considered to be morphologically normal, it refers to the following: i) Head: This is smooth, with a regular contour, generally oval-shaped. It shall be a well-defined acrosomal region that includes 40 - 70% of the head region. The acrosomal region shall not contain large vacuoles, shall not contain more than two small vacuoles, and shall not occupy more than 20% of the sperm head. The post-acrosomal region shall not contain any vacuoles. ii) Mid-piece: This is slender, regular, and approximately the same length as the sperm head. The main axis of the mid-piece shall be aligned with the main axis of the sperm head. Residual cytoplasm is considered abnormal only if it is excessive, i.e., exceeds one-third of the sperm head size. iii) Principal piece: This has a uniform diameter along its length, is thinner than the mid-piece, and is approximately

[0328] μm long (about 10 times the length of the head). It may loop back on itself provided there are no acute angles indicating flagellar breakage.

[0328] Semen samples were collected immediately before the start of treatment (t = 0) and after 3 months of treatment with a dietary supplement of 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 [[END

[0329] Statistical analysis First, descriptive statistics (mean and SEM) were determined for each parameter. Then, a non-parametric Wilcoxon test for paired samples was applied to determine the statistically significant differences in the data recorded before and after T. tuyi supplementation. Significance was accepted for p < 0.05. All analyses were performed using Prism 6 software.

[0330] Results Example 1 Semen volume (VOL) As shown in Figure 1, a highly statistically significant increase in VLO was observed after 3 months of T. tuyi supplementation. The VOL parameter reached an average value of 3.09 ml, representing an increase of nearly 35% with respect to the average value (2.29 ml) observed at the start of the trial.

[0331] When the trial participants were classified with respect to the LRL established by the WHO for this parameter (Figure 2), it was found that 90% of the participants had values higher than the LRL at t = 0, but after trial completion, all recorded values (100%) were higher than the LRL.

[0332] Example 2 Sperm concentration (CONC) A highly statistically significant increase in the CONC parameter was also observed after 3 months of T. tuyi supplementation (Figure 3). In fact, the average value was 17.16×10 6 spermatozoa / ml, representing an increase of approximately 86% with respect to the initial average value (9.22×10 6 spermatozoa / ml) recorded at t = 0. Importantly, such an average value after 3 months of dietary supplementation with T. tuyi was higher (by 14.4%) than the LRL (15×10 6 spermatozoa / ml) established by the WHO.

[0333] In this case, the effect of Tribulus terrestris supplementation was dramatically positive when looking at the individual classifications according to the LRL provided by the WHO. There were no participants who showed values > LRL at t = 0, i.e., 100% presented oligospermia, but this percentage decreased to 50% after 3 months of Tribulus terrestris intake.

[0334] Example 3 Total sperm count (NUM) in ejaculate Figure 5 summarizes the results obtained in the test. A strong significant increase in NUM was detected as a result of Tribulus terrestris supplementation. At t = 0, the average value was 22.20×10 6 sperm per ejaculate, but after 3 months of daily Tribulus terrestris intake, the recorded average value was 52.75×10 6 sperm per ejaculate, representing an increase of approximately 238% with respect to the initial value. Also, regarding the LRL proposed by the WHO for this parameter, the observed changes were dramatic: at t = 0, the average value was significantly lower than the LRL (representing only 57% of this reference value), but after 3 months of Tribulus terrestris intake, the average value was approximately 35% higher than the LRL.

[0335] Excellent results were again observed regarding the distribution of the test participants according to the LRL established by the WHO for the NUM parameter. As shown in Figure 6, only 1 individual (5%) showed values of NUM > LRL, i.e., 95% of them (19 out of 20) presented oligospermia. However, after 3 months of Tribulus terrestris intake, the percentage of participants with oligospermia decreased to 35%.

[0336] Example 4 Progressive motility (PR) As observed with the aforementioned parameters, a statistically significant increase was also detected in PR after 3 months of T. chuii intake (Figure 7). The mean value changed from 26.1% at the start of the trial to 32.85% at the end, representing a net increase of 25.9% in this parameter. Additionally, it must be emphasized that even the final mean value after 3 months of T. chuii supplementation was slightly higher than the LRL (32%) provided by the WHO for PR.

[0337] When dealing with the classification of participants regarding the LRL provided by the WHO for PR (Figure 8), 75% of the participants presented asthenozoospermia at t = 0 (only 25% showed values > LRL), but this percentage decreased to 55% as a result of 3 months of T. chuii supplementation (45% of the participants had values > LRL).

[0338] Example 5 Normal form (MORF) As shown in Figure 9, in contrast to all the aforementioned parameters already described, no statistically significant changes were detected in the recorded mean values at both the start (6.65%) and end (6.45%) of the trial. Also, both mean values were higher than the LRL (4%) provided by the WHO for this parameter.

[0339] However, when we looked at the distribution of participants regarding the LRL, we observed that 35% presented teratozoospermia at t = 0 (i.e., 65% showed values > LRL), but this percentage of individuals with teratozoospermia decreased to 10% (90% of the individuals showed values > LRL) at the end of the trial (Figure 10).

[0340] Example 6 Normal semen According to WHO(2010), individuals with normal semen have a total sperm count (or concentration depending on the reported result) above the LRL, as well as percentages of progressively motile (PR) sperm and morphologically normal sperm (NF). Thus, this state of normal semen is evident in relation to the analysis as it encompasses important parameters of semen quality. As shown in Figure 11, individuals representing normal semen at t = 0 were not included in the study (as described in the introduction, individuals were included in the study if they exhibited oligospermia, asthenospermia, and / or teratozoospermia). However, after 3 months of T. chuii intake, the percentage of participants with normal semen increased up to 30% (6 out of 20 individuals).

[0341] Conclusion Taking into account all the data, the results obtained by supplementation with T. chuii (250 mg / day for 3 months) revealed a statistically significant positive response in the mean values of 4 out of the 5 parameters analyzed in the semen samples. When dealing with the percentage of individuals classified by the LRL (provided by WHO, 2010) in each of these parameters, positive results were obtained in all cases. Thus, administration of the microalgae T. chuii improves semen quality in infertile men with abnormal semen parameters.

Claims

1. A pharmaceutical composition comprising the biomass of Tetraselmis chuii (T. chuii) for use in the treatment of infertility in male patients.

2. The pharmaceutical composition according to claim 1, wherein the patient has oligospermia.

3. The pharmaceutical composition according to any one of claims 1 or 2, wherein the total number and / or concentration of spermatozoa in a semen sample of the patient after the treatment is equal to or higher than a reference value.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the patient has asthenospermia.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the percentage of progressively motile spermatozoa in a semen sample of the patient after the treatment is equal to or higher than a reference value.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the patient has teratozoospermia.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the percentage of morphologically normal spermatozoa in a semen sample of the patient after the treatment is equal to or higher than a reference value.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the patient has hypospermia.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the volume of semen per ejaculate in a semen sample of the male patient after the treatment is equal to or higher than a threshold reference.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the biomass of T. chuii is dehydrated.

11. The pharmaceutical composition according to claim 1, wherein the treatment comprises administering 10 to 500 mg / day, preferably 250 mg / day, of the dehydrated biomass.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the treatment comprises administering the pharmaceutical composition for at least one month.

13. A pharmaceutical composition comprising the biomass of T. chuii for improving the quality of semen of male subjects having normal semen, not having hypospermia, and not having a disorder characterized by a percentage of SDF above a reference value.

14. An oral supplement comprising the biomass of T. chuii for improving the quality of semen of male subjects having normal semen, not having hypospermia, and not having a disorder characterized by a percentage of SDF above a reference value.

15. The pharmaceutical composition according to claim 13 or the oral supplement according to claim 14, wherein the biomass of T. thuisii is dehydrated.

16. The pharmaceutical composition according to claim 13 or the oral supplement according to claim 14, wherein the biomass of T. thuisii is administered as dehydrated biomass at 10 to 500 mg / day, preferably 250 mg / day.

17. The pharmaceutical composition according to claim 13 or the oral supplement according to claim 14, wherein the pharmaceutical composition or oral supplement is administered for at least one month.

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