Novel composition and use thereof in improving sperm cell viability, survival rate and longevity as well as fertility

Abstract

The present invention relates to novel compositions, methods of making the novel compositions, and their use in increasing the viability, survival and longevity and fertility of sperm cells.

Description

TECHNICAL FIELD

[0001] The present invention relates to a novel composition, a method of manufacturing said composition and its use in improving the motility, viability and longevity of sperm cells and fertility. BACKGROUND

[0002] One in six couples suffers from infertility, and one in two cases is due to male defects.

[0003] Depending on the severity of male infertility, various methods can be used to help couples, such as artificial insemination (AI) for mild sperm dysfunction, in particular intrauterine insemination (IUI), in vitro fertilization (IVF) for moderate sperm dysfunction and intra-cytoplasmic sperm injection (ICSI) for males with severe sperm dysfunction.

[0004] In this assisted reproduction technology (ART), freezing of sperm samples is usually necessary.

[0005] However, one major drawback of frozen sperm samples is that once thawed, the motility, viability and longevity of previously frozen sperm are reduced compared to fresh sperm samples, and the number of progressively motile sperm is also reduced.

[0006] Another drawback of the freezing process is that it accelerates the maturation process of sperm, i.e. capacitation. Capacitation is a necessary step in sperm maturation that usually occurs in the female reproductive tract. Although capacitation is a necessary step in sperm maturation, it causes irreversible changes in sperm and ultimately impairs semen quality, motility and fertilization capacity. Capacitation causes a sharp increase in sperm metabolic activity and shortens the total longevity of sperm in the female reproductive tract. This impairment is particularly severe when insemination is not synchronized with ovulation.

[0007] In addition, for farm animals that use artificial insemination (AI) extensively, the decrease in frozen sperm motility means that the concentration of sperm in the pipette must be increased, which reduces the overall success rate of the method.

[0008] To avoid the drawbacks associated with the freezing process, it has been found that the use of a non-toxic agent can improve sperm motility, viability and longevity, thereby improving the pregnancy rate in ART methods, thus showing strong therapeutic value in medically assisted reproduction, not only for human treatment but also for the industrial reproduction of many species, particularly animal species.

[0009] In particular, it was surprisingly found that Slo3 potassium channel inhibitors can be used to increase sperm motility, viability and longevity, thereby increasing the pregnancy rate in ART methods.

[0010] However, these agents can not be in sufficient contact with the sperm, thereby affecting their activity, and the sperm sample can also suffer physical damage, such as sperm cell sedimentation.

[0011] Therefore, there is a need to provide a new non-toxic sperm composition for ART methods, in order to enhance the action of agents such as Slo3 potassium channel inhibitors when used (thereby also reducing the amount of agents such as Slo3 potassium channel inhibitors), to gradually release the sperm over time, in particular to ensure a longer diffusion of the sperm cells in the uterus during artificial insemination, while also avoiding physical damage such as sperm cell sedimentation.

[0012] Surprisingly and unexpectedly, the inventors have found a new non-toxic composition which can increase the contact time between agents such as Slo3 potassium channel inhibitors and sperm cells (without negatively affecting their activity), gradually release the sperm over time, in particular to ensure a longer diffusion of the sperm cells in the uterus during artificial insemination (thereby extending the insemination window by extending the fertilizing capacity of the sperm), and can also avoid physical damage such as sperm cell sedimentation. SUMMARY

[0013] The present invention relates to a composition comprising:

[0014] - human and / or animal semen comprising sperm cells, such as sperm,

[0015] - a compound selected from the group consisting of collagen peptides, polymers and copolymers comprising polysaccharides, and mixtures thereof,

[0016] - a semen diluent,

[0017] - optionally, a Slo3 potassium channel inhibitor, and

[0018] - optionally, a lipid, such as a liposome.

[0019] The present invention also relates to a gel comprising the composition as defined previously.

[0020] The present invention also relates to a kit comprising the composition as defined previously.

[0021] The present invention also relates to a method of manufacturing the composition as defined previously or the gel as defined previously, comprising the following steps:

[0022] - mixing a semen diluent with a compound, optionally a metal ion, a Slo3 potassium channel inhibitor and / or a lipid substance (e.g. a liposome), and optionally additional compounds and / or divalent or trivalent ion chelators, to obtain a mixture,

[0023] - diluting said mixture with water to obtain a diluted mixture, and

[0024] - mixing human and / or animal semen into said diluted mixture.

[0025] The present application also relates to the use of a composition as defined previously or of a gel as defined previously for increasing the contact time between sperm cells and a Slo3 potassium channel inhibitor.

[0026] The present application also relates to the use of a composition as defined previously or of a gel as defined previously for preventing the sedimentation of sperm cells.

[0027] The present application also relates to a composition as defined previously or to a gel as defined previously for use in improving the motility, viability and longevity of sperm cells, in particular spermatozoa, preferably capacitated spermatozoa.

[0028] The present application also relates to a composition as defined previously or to a gel as defined previously for use in improving fertility. DETAILED DESCRIPTION

[0029] For the purposes of the present application, the numerical ranges are ranges including the endpoints, unless otherwise stated.

[0030] The present application relates to a composition comprising:

[0031] - human and / or animal semen comprising sperm cells (e.g. spermatozoa),

[0032] - a compound selected from the group consisting of collagen peptides, polymers and copolymers (including polysaccharides) and mixtures thereof,

[0033] - a semen diluent,

[0034] - optionally, a Slo3 potassium channel inhibitor, and

[0035] - optionally, a lipid, e.g. a liposome.

[0036] The inventors have surprisingly found that the aforementioned composition is non-toxic, in particular has an anti-crystallization effect during freezing (by inhibiting the devitrification of spermatozoa during freezing), is able to prevent the oxidation of sperm cells, maintains the integrity of the acrosome of sperm cells, improves the mitochondrial activity of sperm cells, while improving the motility, viability and longevity of sperm cells, thereby improving fertility.

[0037] The sperm cells can be fresh sperm cells and in particular freshly ejaculated sperm cells, frozen sperm cells, thawed sperm cells and / or sperm cells recovered from the epididymis or testis.

[0038] "Sperm cells" are preferably understood as spermatozoa.

[0039] "Semen diluent" is preferably understood as any liquid diluent, preferably an aqueous solution, used to increase the volume of semen up to the desired dose while preserving the functional properties of the sperm cells, in particular their fertilizing capacity.

[0040] Examples of semen diluents include, but are not limited to, media comprising one or more of the following compounds: sugars (e.g. glucose, galactose, fructose, ribose, trehalose and mixtures thereof), buffers (e.g. citrate (in particular sodium citrate), phosphate, bicarbonate (in particular sodium bicarbonate), TES (N-[tris(hydroxymethyl)methyl]-2- aminethanesulfonic acid), HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)), MOPS (3-(N-morpholino)propanesulfonic acid), Tris (tris(hydroxymethyl)aminomethane) and mixtures thereof), inorganic ion salts (e.g. sodium chloride, potassium chloride, sodium tartrate, potassium tartrate, sodium acetate, potassium acetate, sodium sulfate, potassium sulfate and mixtures thereof), protein-based compounds (e.g. peptone, milk, lactose, egg yolk, egg phosphatidylcholine (EPC), bovine serum albumin (BSA) and mixtures thereof), chelating agents such as EDTA (ethylenediaminetetraacetic acid), amino acids such as cysteine, amino acid-based compounds such as acetylcysteine, antibiotics such as penicillin and streptomycin, polyvinyl alcohol (PVA), glycerol, detergents, citric acid and mixtures thereof.

[0041] In particular, examples of semen diluents can be as described in the literature: Semen extenders used in the artificial insemination of swine, Gadea et al., SPANISH JOURNAL OF AGRICULTURAL RESEARCH 1 (2): 17-27 and / or Liposomes for cryopreservation of bovine sperm, T et al., Theriogenology, 2011, Nov; 76(8): 1465-72.

[0042] Preferably, examples of semen diluents include, but are not limited to: Beltsville Liquid (BL-1), Beltsville Thawing Solution (BTS), Illinois Variable Temperature (IVT), Kiev, plus, Modena, MULBERRY Reading, Zorlesco, ZORPVA, SpermAid, Safe Cell Plus, BF-5, BW25, TCF medium, NutriXcell ultra, NutriXcell plus, OptiXcell, OptiXcell Alpha, and mixtures thereof. More preferably, the semen diluent is selected from OptiXcell, OptiXcell Alpha, and mixtures thereof.

[0043] Polymers and copolymers including polysaccharides can be anionic polymers and copolymers (including anionic polysaccharides), cationic polymers and copolymers (including cationic polysaccharides), non-ionic polymers and copolymers (including non-ionic polysaccharides), covalently grafted polymers and copolymers (e.g., polymers and copolymers grafted with amine functional groups), thermally responsive (or temperature responsive) polymers and copolymers (e.g., LCST polymers and copolymers and UCST polymers and copolymers), including polymers and copolymers grafted from, onto, or through polysaccharides, and mixtures thereof.

[0044] Examples of anionic polymers and copolymers include, but are not limited to, anionic polysaccharides, such as anionic gums, in particular xanthan gum, gum arabic, carrageenan, arabinogalactan gum, gellan gum, and mixtures thereof, alginates, pectin, hyaluronic acid (HA), cellulose derivatives (e.g., carboxymethylcellulose (CMC)), proteins (e.g., gelatin and egg white proteins), and mixtures thereof.

[0045] Examples of cationic polymers and copolymers include, but are not limited to, cationic polysaccharides (e.g., chitosan), cationic derivatives or cationically modified polysaccharides of polysaccharides (e.g., cellulose, dextran, and guar gum), proteins (e.g., gelatin and egg white proteins), and mixtures thereof.

[0046] Examples of non-ionic polymers and copolymers include, but are not limited to, non-ionic polysaccharides, in particular microcrystalline cellulose (MCC), cellulose derivatives (e.g. methylcellulose, hydroxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose (HPMC) and mixtures thereof), scleroglucan, pullulan, ethylene glycol methacrylate polymers (PEGMA), triethylene glycol methacrylate polymers (PTEGMA), oligoethylene glycol methyl ether methacrylate polymers (POEGMA), poly(vinyl methyl ether) (PVME), poly(N-isopropylacrylamide) (PNiPAM) and copolymers thereof, poly(N-vinylcaprolactam) (PNVCL), poloxamers such as F-127 (CAS 9003-11-6), poly(ethylene oxide-co-propylene oxide) (PEPO), polyoxazolines and mixtures thereof.

[0047] Examples of covalently grafted polymers and copolymers (e.g. polymers and copolymers grafted with amine functional groups) include, but are not limited to, homopolymers and copolymers of N-substituted poly(acrylamides), homopolymers and copolymers of poly(methacrylamides), acrylate polymers and copolymers (e.g. methacrylate polymers and copolymers), acrylamide polymers and copolymers (e.g. methacrylamide polymers and copolymers), LCST polymers and copolymers and UCST polymers and copolymers, amino-terminated poloxamers, amino-terminated poly(N-isopropylacrylamide) (PNiPAM), e.g. amino-terminated poly(N-isopropylacrylamide) (PNiPAM) using cysteamine, amino-terminated poly(ethylene oxide-co-propylene oxide) (PEPO), amino-terminated poly(N-vinylcaprolactam) (PNVCL), amino-terminated ethylene glycol methacrylate polymers (PEGMA), amino-terminated polyoligo(ethylene glycol) methyl ether methacrylate (POEGMA), amino-terminated polyoxazolines and mixtures thereof.

[0048] Examples of thermoresponsive polymers (or temperature-responsive polymers) (e.g. LCST polymers and copolymers and UCST polymers and copolymers) include, but are not limited to: LCST polymers and copolymers, UCST polymers and copolymers, polymers and / or copolymers as described before grafted to a polysaccharide (backbone) as defined by the present invention (side chains), preferably polymers and copolymers grafted with hyaluronic acid (HA) and / or alginate (Alg), in particular sodium alginate, such as HA-g-PEPO, Alg-g-PEPO, HA-g-PNiPAm, Alg-g-PNiPAm, HA-g-PEGMA, Alg-g-PEGMA, HA-g-PTEGMA, Alg-g-PTEGMA, HA-PEPO, Alg-PEPO, HA-PNiPAm, Alg-PNiPAm, HA-PEGMA, Alg-PEGMA, HA-PTEGMA, Alg-PTEGMA, and mixtures thereof, more preferably HA-g-PNiPAm and mixtures thereof.

[0049] Examples of LCST polymers include, but are not limited to: N-substituted poly(acrylamides) and poly(methacrylamides), poly(N-vinylamides), protein-related polymers, poly(methyl 2-alkylamidopropionates), poly(oxazolines), poly(oxides), poly(vinyl ethers), poly[oligoethylene glycol (meth)acrylates], poly(phosphoesters), alkyl celluloses (e.g. methyl cellulose and hydroxypropyl methyl cellulose), and mixtures thereof.

[0050] Examples of UCST polymers include, but are not limited to: poly(sulfobetaines), N-substituted poly(acrylamides) and poly(methacrylamides) (e.g. poly(N-acrylglycamide), poly(N-cyanomethylacrylamide)), poly(acrylamides) and copolymers (e.g. poly(acrylamide-co-acrylonitrile)), poly(N-acrylasparagine), and mixtures thereof.

[0051] Preferably, the backbone of the polymer has a molar mass between 10000 g / mol and 2000000 g / mol, preferably between 50000 g / mol and 1000000 g / mol, preferably between 50000 g / mol and 400000 g / mol, more preferably between 100000 g / mol and 300000 g / mol, even more preferably between 150000 and 250000 g / mol.

[0052] Preferably, the side chains of the polymer have a chain molar mass between 500 g / mol and 70000 g / mol, preferably between 2000 g / mol and 50000 g / mol, more preferably between 2000 g / mol and 40000 g / mol.

[0053] Preferably, the ratio of polymer side chain to polymer main chain is between 0:100 and 100:0, more preferably between 20:80 and 80:20, and even more preferably between 40:60 and 80:20.

[0054] Preferably, the concentration of the polymer or copolymer comprising the polysaccharide is from 0.25% to 4% by weight relative to the total weight of the composition, more preferably from 0.4% to 3% by weight, and more preferably from 0.8% to 2.5% by weight.

[0055] Preferably, the polymers and copolymers as defined above are biocompatible polymers and copolymers.

[0056] "Biocompatible polymers or copolymers" are preferably understood as polymers and / or copolymers that are suitable for exposure to the body and bodily fluids and that can improve bodily functions without altering their normal function and without causing allergies or other side effects.

[0057] Preferably, the polymer and copolymer are selected from the following substances: polysaccharides, gelatin, ethylene glycol methacrylate polymer (PEGMA), triethylene glycol methacrylate polymer (PTEGMA), polyoligo(ethylene glycol) methyl ether methacrylate polymer (POEGMA), N-isopropylacrylamide polymer (PNiPAM), poly-N-vinylcaprolactam polymer (PNVCL), poloxamer, etc. F-127 (CAS 9003-11-6), poly(ethylene oxide-co-propylene oxide) polymer (PEPO), covalently grafted polymers and copolymers as defined above, such as polymers and copolymers grafted with amino functional groups as defined above, including amino-terminated poloxamer, amino-terminated poly(N-isopropylacrylamide) (PNiPAM), such as amino-terminated poly(N-isopropylacrylamide) (PNiPAM) using cysteine, amino-terminated poly(ethylene oxide-co-propylene oxide) (PEPO), amino-terminated poly(N-vinylcaprolactam) (PNVCL), amino-terminated ethylene glycol methacrylate polymer (PEGMA), amino-terminated polyoligo(ethylene glycol) methyl ether methacrylate polymer (POEGMA), amino-terminated polyoxazoline and mixtures thereof, such as compounds grafted from polysaccharides, grafted onto polysaccharides or grafted through polysaccharides, as defined above.

[0058] Examples of polysaccharides according to the present invention include, but are not limited to, gums (e.g., guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum, gellan gum, and mixtures thereof), cellulose (e.g., microcrystalline cellulose (MCC) and cellulose derivatives (e.g., methylcellulose, carboxymethylcellulose (CMC), hydroxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose (HPMC), and mixtures thereof), alginates (e.g., sodium alginate), pectin, pullulan, chitosan, hyaluronic acid (HA), dextran, sclerotium polysaccharides, and mixtures thereof).

[0059] Pectin can be selected from low molecular weight (LM) pectin, preferably citrus pectin, apple pectin and mixtures thereof.

[0060] Examples of polysaccharide mixtures include, but are not limited to, mixtures of pectin and carboxymethyl cellulose (CMC) or mixtures of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), and mixtures thereof.

[0061] Examples of collagen peptides include, but are not limited to, peptan.

[0062] Preferably, the concentration of collagen peptides is from 0.1% to 10% by weight relative to the total weight of the composition, more preferably from 0.2% to 5% by weight, and more preferably from 0.5% to 2.5% by weight.

[0063] The composition may also contain metal ions, preferably divalent and / or trivalent metal ions, more preferably selected from beryllium ions, magnesium ions, calcium ions, strontium ions, barium ions, radium ions and mixtures thereof.

[0064] Metal ions can be in the form of inorganic salts, such as chlorides, especially beryllium chloride (BeCl2), magnesium chloride (MgCl2), calcium chloride (CaCl2), strontium chloride (SrCl2), barium chloride (BaCl2), radium chloride (RaCl2), and mixtures thereof.

[0065] The concentration of the metal ions in the composition as defined above can be in the range of 0.2 mM to 1 M, preferably 0.5 mM to 80 mM, and more preferably 1 mM to 50 mM.

[0066] The concentration of the compound can be from 0.001% to 10% by weight relative to the total weight of the composition, preferably from 0.01% to 10% by weight, and more preferably from 0.1% to 5% by weight.

[0067] The composition may also contain one or more additional compounds selected from the following:

[0068] - Sialic acid, and / or

[0069] - Sugars, such as mannose, glucosamine and their derivatives, and mixtures thereof, and / or

[0070] - Oligosaccharides, such as oligomannose, and / or

[0071] - A binder with affinity for sialic acid, and / or

[0072] - A binding agent with an affinity for sugars (e.g., mannose, glucosamine and their derivatives and mixtures thereof), and / or

[0073] - A binding agent with affinity for oligosaccharides (e.g., oligomannose), and / or

[0074] - A reagent, preferably an enzyme, for hydrolyzing sialic acid, and / or

[0075] - A reagent, preferably an enzyme, for hydrolyzing sugars (e.g., mannose, glucosamine and their derivatives, and mixtures thereof), and / or

[0076] - A reagent for hydrolyzing oligosaccharides (e.g., oligomannose), preferably an enzyme.

[0077] Examples of glucosamine derivatives include, but are not limited to, N-acetamidoglucose (Glc-NAc), β-D(1,3)-galactosamine, and mixtures thereof.

[0078] Examples of binders with affinity for sialic acid include, but are not limited to, compounds described in patent application WO 2017012993(A1), particularly monoclonal antibodies, lectins that interact with sialic acid, such as elderberry lectin (Sambucus NigraAgglutinin, SNA), wheat lectin (Triticum Vulgaris Lectin, WGA), snowdrop lectin or lectin (Galanthus Nivalis Lectin / Agglutinin, GNL / GNA), ovalbumin, novel glycoproteins, and mixtures thereof.

[0079] Examples of binders having an affinity for sugars (such as mannose, glucosamine and their derivatives and mixtures thereof) and / or an affinity for oligosaccharides (such as oligomannose) include, but are not limited to, lectins, antibodies, boric acid and mixtures thereof.

[0080] Examples of enzymes that hydrolyze sialic acid include, but are not limited to, sialidase.

[0081] Examples of enzymes that hydrolyze sugars (such as mannose, glucosamine and their derivatives) include, but are not limited to, neuraminidase.

[0082] Examples of enzymes that hydrolyze oligosaccharides (e.g., oligomannose) include, but are not limited to, compounds described in patent application WO 2017012993 (A1), particularly transglucosidases, hydrolase compounds, and mixtures thereof.

[0083] Preferably, these additional compounds, particularly lectins, sialidases, and antibodies, are natural molecules, preferably unmodified molecules, and more preferably unmodified molecules without polyethylene glycol (PEG) groups.

[0084] Examples of lectins include, but are not limited to, wheat germ agglutinin (WGA).

[0085] The concentration range of the additional compound can be from 0.001 mg / L to 1000 mg / L, preferably from 0.01 mg / L to 100 mg / L, and more preferably from 0.02 mg / L to 50 mg / L.

[0086] For example, when the added compound is a lectin, its concentration range can be from 0.001 mg / L to 100 mg / L, preferably from 0.01 mg / L to 50 mg / L, and more preferably from 0.02 mg / L to 10 mg / L.

[0087] For example, when the additional compound is sialic acid, its concentration range can be from 0.1 mg / L to 1000 mg / L, preferably from 1 mg / L to 100 mg / L, and more preferably from 10 mg / L to 50 mg / L.

[0088] Preferably, the binder is a mixture of sialic acid and lectin (e.g., wheat germ lectin (WGA)).

[0089] Preferably, the composition comprises:

[0090] 0.001 mg / L to 100 mg / L, preferably 0.01 mg / L to 50 mg / L, more preferably 0.02 mg / L to 10 mg / L of lectin, and

[0091] Sialic acid ranging from -0.1 mg / L to 1000 mg / L, preferably from 1 mg / L to 100 mg / L, and more preferably from 10 mg / L to 50 mg / L.

[0092] Preferably, the composition comprises:

[0093] - Human and / or animal semen containing sperm cells (such as sperm),

[0094] - A compound selected from collagen peptides, polymers and copolymers (including polysaccharides) as defined above, and mixtures thereof, preferably sclerotium polysaccharides, microcrystalline cellulose (MCC), carboxymethyl cellulose (CMC), mixtures of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), HA-g-PNIPAm, Alg-g-PNIPAm, combinations of pectin and metal ions (e.g., strontium ions), combinations of alginate and metal ions (e.g., barium ions), and mixtures thereof, more preferably mixtures of sclerotium polysaccharides, microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), combinations of pectin and strontium ions, HA-g-PNIPAm, and mixtures thereof.

[0095] - Semen diluents as defined above, preferably NutriXcell ultra, NutriXcell plus, OptiXcell, OptiXcell Alpha, and mixtures thereof.

[0096] -Optionally, Slo3 potassium channel inhibitors

[0097] -Optionally, lipids, such as liposomes, and

[0098] -Optionally, additional compounds as defined above, and preferably a mixture of sialic acid and lectin, such as wheat germ lectin (WGA).

[0099] Anti-settling effect

[0100] The inventors unexpectedly discovered that the aforementioned composition can be used to prevent sperm cell sedimentation (anti-settling effect), particularly by forming a weak network capable of capturing sperm cells during storage. Preferably, compounds selected from collagen peptides, polymers, and copolymers (including polysaccharides) and mixtures thereof form a matrix with the semen diluent capable of capturing sperm cells during storage. Advantageously, compounds selected from collagen peptides, polymers, and copolymers (including polysaccharides) and mixtures thereof are not used as coating or embedding materials, but rather form a matrix with the semen diluent to capture sperm cells during storage.

[0101] When the composition is used to prevent sperm cell precipitation, the concentration of the compound is preferably 0.001% to 5% by weight of the total weight of the composition, more preferably 0.01% to 4% by weight, and even more preferably 0.1% to 3% by weight.

[0102] When the composition is used to prevent sperm cell deposition, the compound is preferably a polysaccharide as defined above, more preferably selected from the following substances: gelatin, microcrystalline cellulose (MCC), methylcellulose, carboxymethyl cellulose (CMC), hydroxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose (HPMC), pullulan, dextran, gums (e.g., guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum and mixtures thereof), sclerotium polysaccharide, peptan and mixtures thereof.

[0103] More preferably, when the composition is used to prevent sperm cell sedimentation, the compound is selected from sclerotium polysaccharides, microcrystalline cellulose (MCC), carboxymethyl cellulose (CMC), and mixtures thereof, such as a mixture of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC).

[0104] Preferably, the composition comprises:

[0105] - Human and / or animal semen containing sperm cells (such as sperm),

[0106] - A compound selected from the following substances: collagen peptides, polymers and copolymers (including polysaccharides) as defined above, and mixtures thereof, preferably sclerotium polysaccharides, microcrystalline cellulose (MCC), carboxymethyl cellulose (CMC), mixtures of microcrystalline cellulose (MCC) and carboxymethyl cellulose (CMC), and mixtures thereof.

[0107] - Semen diluents as defined above, preferably NutriXcell ultra, NutriXcell plus, OptiXcell, OptiXcell Alpha, and mixtures thereof.

[0108] -Optionally, Slo3 potassium channel inhibitors

[0109] -Optionally, lipids, such as liposomes, and

[0110] - Optionally, additional compounds as defined above, and preferably a mixture of sialic acid and lectin, such as wheat germ lectin (WGA).

[0111] Gel effect

[0112] The inventors unexpectedly discovered that the compositions of the present invention can form a gel in which the present Slo3 potassium channel inhibitor is retained, thereby increasing the contact time between the Slo3 potassium channel inhibitor and sperm cells and enhancing its effect on sperm cells, especially during artificial insemination and intrauterine dilution. Advantageously, the time of action of the inhibitor from the start of artificial insemination (AI) to the arrival of sperm cells near the oocyte is prolonged, thereby prolonging the fertilization time of the artificial insemination dose. Advantageously, compounds selected from collagen peptides, polymers and copolymers (including polysaccharides) and mixtures thereof are not used as coating or embedding materials, but rather form a gel with the semen diluent.

[0113] The inventors also unexpectedly discovered that the composition of the present invention can gradually release sperm over time, particularly ensuring that sperm cells diffuse within the uterus for a longer period during artificial insemination (thereby increasing the fertilization window by maintaining sperm fertility for a longer period).

[0114] Advantageously, the composition described above can form a gel that is non-toxic to sperm cells and also non-toxic to the uterus (and has no physiological effects), which can retain sperm cells and release viable sperm cells over time during procedures such as artificial insemination, in vitro fertilization (IVF), and / or intracytoplasmic sperm injection (ICSI).

[0115] When the composition is used to form a gel, the concentration of the compound is preferably 0.1% to 10% by weight of the total weight of the composition, more preferably 0.5% to 5% by weight, and more preferably 1% to 3% by weight.

[0116] When the composition is used to form a gel, the compound is preferably selected from methylcellulose, carboxymethylcellulose (CMC), hydroxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose (HPMC), alginate (Alg), pectin, pullulan, chitosan, hyaluronic acid (HA), dextran, gums (e.g., guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum and mixtures thereof), peptan, polymers and copolymers (e.g., ethylene glycol methacrylate polymer (PEGMA), triethylene glycol methacrylate polymer (PTEGMA), polyoligo(ethylene glycol) methyl ether methacrylate (POEGMA), N-isopropylacrylamide polymer (PNiPAM), poly-N-vinylcaprolactam (PNVCL), poloxamer (e.g.) F-127 (CAS 9003-11-6)), poly(ethylene oxide-co-propylene oxide) polymer (PEPO), and mixtures thereof, such as HA-g-PNIPAm and Alg-g-PNIPAm, more preferably HA-g-PNIPAm, Alg-g-PNIPAm, and mixtures thereof.

[0117] Preferably, the compound is selected from carboxymethyl cellulose (CMC), alginate, pectin, chitosan, hyaluronic acid (HA), gums (such as guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum) and mixtures thereof, and is combined with metal ions, preferably selected from beryllium ions, magnesium ions, calcium ions, strontium ions, barium ions, radium ions and mixtures thereof, more preferably a combination of pectin and metal ions such as strontium ions, or a combination of alginate and metal ions such as barium ions.

[0118] Preferably, the composition comprises:

[0119] - Human and / or animal semen containing sperm cells (such as sperm),

[0120] - A compound selected from the following substances: collagen peptides, polymers and copolymers (including polysaccharides) as defined above, and mixtures thereof, preferably HA-g-PNIPAm, Alg-g-PNIPAm, combinations of pectin and metal ions (e.g., strontium ions), combinations of alginate and metal ions (e.g., barium ions), and mixtures thereof, more preferably HA-g-PNIPAm, Alg-g-PNIPAm, combinations of pectin and strontium ions, and mixtures thereof.

[0121] - Semen diluents as defined above, preferably NutriXcell ultra, NutriXcell plus, OptiXcell, OptiXcell Alpha, and mixtures thereof.

[0122] -Optionally, Slo3 potassium channel inhibitors

[0123] -Optionally, lipids, such as liposomes, and

[0124] - Optionally, additional compounds as defined above, and preferably a mixture of sialic acid and lectin, such as wheat germ lectin (WGA).

[0125] Slo3 potassium channel inhibitors can be selected from barium, mibeladin, chlorfenapyr, quinidine, and mixtures thereof.

[0126] The concentration range of the Slo3 potassium channel inhibitor can be from 0.01 μM to 10 μM, preferably from 0.05 μM to 5 μM, and more preferably from 0.1 μM to 2 μM.

[0127] Examples of lipids include, but are not limited to, liposomes, such as liposomes derived from egg yolk phospholipids and liposomes derived from soybean lecithin, and mixtures thereof.

[0128] The concentration of lipid substances, such as liposomes, can be from 0.01% to 10% of the total weight of the composition, preferably from 0.05% to 5% of the total weight, and more preferably from 0.1% to 1% of the total weight.

[0129] Animal semen may be selected from mammalian semen, such as bull, bovine, pig, horse, goat, sheep, buffalo, camel, aquatic animal (e.g., fish, crustaceans and mollusks), insect, cat and / or dog semen, and avian semen, such as chicken and / or turkey semen, preferably from avian, bovine and / or pig semen, more preferably from bovine and / or pig semen.

[0130] The compositions as defined above may also contain divalent or trivalent ion chelating agents, preferably photolytic divalent or trivalent ion chelating agents, more preferably selected from o-nitrobenzyl compounds, such as o-nitrobenzyl-BAPTA and DMNP-EDTA, and mixtures thereof.

[0131] In particular, the inventors have unexpectedly discovered that the use of divalent or trivalent ion chelating agents, especially photolytic divalent or trivalent ion chelating agents, can chelate divalent or trivalent ions, thereby preventing and / or limiting the gelation process and / or increasing the viscosity of the composition. Then, when the composition is subjected to external stimuli (e.g., light within a given wavelength range, pH value, temperature, ionic forces, and combinations thereof), the chelated divalent or trivalent ions are released, the aforementioned compounds crosslink, forming a gel and / or increasing viscosity. This can trap sperm cells, thereby ensuring the controlled release of sperm cells during fertilization, preferably within 72 hours, more preferably between 24 and 48 hours, while also increasing the contact time between sperm cells and Slo3 potassium channel inhibitors (if used).

[0132] Advantageously, the divalent or trivalent ion chelating agents as defined above are non-toxic to semen, especially sperm cells, and also non-toxic to the uterus (and have no physiological effects), have antioxidant and acrosome protective properties, and can participate in triggering thickening properties and anti-settling effects, or can be used as adjuvants in gelling formulations for the controlled release of sperm cells, preferably within 72 hours, more preferably within 24 to 48 hours.

[0133] The concentration of the chelating agent can range from 0.005% to 5% relative to the total weight of the composition, preferably from 0.01% to 3% relative to the total weight, and more preferably from 0.05% to 1% relative to the total weight of the composition.

[0134] The ratio of semen to chelating agent can be between 10 and 200, preferably between 20 and 180, and more preferably between 50 and 150.

[0135] Preferably, the dosage of sperm cells contained in the composition is significantly lower than that used in conventional artificial insemination. More preferably, the dosage is at least 5 times lower than that used in conventional artificial insemination, preferably at least 10 times lower, more preferably at least 20 times lower, more preferably at least 30 times lower, more preferably at least 50 times lower, and even more preferably at least 100 times lower.

[0136] The dosage used in conventional artificial insemination typically contains approximately 200 million to 3 billion sperm cells, preferably 2 million to 100 million sperm cells.

[0137] Specifically, when the sperm is selected from bovine sperm, the dose may contain approximately 2 million to 100 million bovine sperm cells.

[0138] Specifically, when the sperm is selected from pig sperm, the dose may contain approximately 200 million to 3 billion pig sperm cells.

[0139] The dosage can range from 0.25 mL to 100 mL.

[0140] For example, when the sperm is selected from bovine sperm, the dosage may include a volume of 0.25 mL to 0.5 mL.

[0141] For example, when the sperm is selected from pig sperm, the dosage may include a volume of 10 mL to 100 mL.

[0142] Preferably, the composition includes a dose of sperm cells between 1 million and 300 million, more preferably between 2 million and 250 million.

[0143] Specifically, when the sperm is selected from bovine sperm, the dose may contain approximately 2 million sperm cells.

[0144] Specifically, when the sperm is selected from pig sperm, the dose may contain approximately 250 million sperm cells.

[0145] The present invention also relates to gels comprising compositions as defined above.

[0146] When the composition is used to form a gel, the composition may have an elastic modulus between 100 Pa and 10000 Pa.

[0147] When the composition is used to prevent sperm cell precipitation, the composition may have a viscosity between 1 and 10,000 mPa·s.

[0148] Preferably, the gel is a thermally responsive gel (or a temperature-responsive gel).

[0149] When the gel is a thermoresponsive gel (or temperature-responsive gel) or when the composition is used to form a thermoresponsive gel (or temperature-responsive gel), the compound as defined above is preferably a polymer and / or copolymer (chain) as defined above, which is grafted onto a polysaccharide (main chain) as defined above.

[0150] The composition may also contain water.

[0151] The present invention also relates to kits comprising compositions as defined above.

[0152] The present invention also relates to a method for manufacturing a composition or gel as defined above, comprising the following steps:

[0153] - The semen diluent is mixed with a compound, optionally a metal ion, an Slo3 potassium channel inhibitor, and / or a lipid (e.g., liposomes), and optionally additional compounds and / or divalent or trivalent ion chelating agents, to obtain a mixture.

[0154] - Dilute the mixture with water to obtain a diluted mixture, and

[0155] - Mix human and / or animal semen into the diluted mixture.

[0156] The present invention also relates to the use of compositions or gels as defined above in increasing the contact time between sperm cells and Slo3 potassium channel inhibitors.

[0157] The present invention also relates to compositions or gels as defined above for increasing the contact time between sperm cells and Slo3 potassium channel inhibitors.

[0158] The present invention also relates to the use of compositions or gels as defined above in the manufacture of medicaments for increasing the contact time between sperm cells and Slo3 potassium channel inhibitors.

[0159] The present invention also relates to a method for increasing the contact time between sperm cells and an Slo3 potassium channel inhibitor, comprising contacting the sperm cells with a composition as defined above or a gel as defined above.

[0160] The present invention also relates to the use of compositions or gels as defined above for preventing sperm cell precipitation.

[0161] The present invention also relates to compositions or gels as defined above for preventing sperm cell precipitation.

[0162] The present invention also relates to the use of the compositions or gels as defined above in the manufacture of medicaments for preventing sperm cell precipitation.

[0163] The present invention also relates to a method for preventing sperm cell precipitation, comprising contacting sperm cells with a composition as defined above or a gel as defined above.

[0164] The present invention also relates to the use of the compositions as defined above in inhibiting sperm depermeability during freezing, preventing sperm cell oxidation, maintaining the integrity of the sperm cell acrosome, and improving sperm cell mitochondrial activity.

[0165] The present invention also relates to the use of compositions or gels as defined above in improving the motility, viability and lifespan of sperm cells, particularly sperm, preferably capacitated sperm.

[0166] The present invention also relates to compositions or gels as defined above for improving the motility, survival rate and lifespan of sperm cells, particularly sperm, preferably capacitated sperm.

[0167] The present invention also relates to the use of compositions or gels as defined above in the manufacture of medicaments for improving the motility, viability and lifespan of sperm cells, particularly sperm, preferably capacitated sperm.

[0168] The present invention also relates to a method for improving the motility, viability and lifespan of sperm cells, particularly sperm, preferably capacitated sperm, comprising contacting the sperm cells with a composition as defined above or a gel as defined above.

[0169] The present invention also relates to the use of compositions or gels as defined above for enhancing fertility.

[0170] The present invention also relates to compositions or gels as defined above for improving fertility.

[0171] The present invention also relates to the use of the compositions or gels as defined above in the manufacture of medicaments for improving fertility.

[0172] The present invention also relates to a method for improving fertility, comprising contacting sperm cells with a composition or a gel as defined above.

[0173] The present invention also relates to the use of compositions or gels as defined above in the treatment of infertility, particularly male infertility.

[0174] The present invention also relates to compositions or gels as defined above for the treatment of infertility, particularly male infertility.

[0175] The present invention also relates to the use of the compositions or gels as defined above in the manufacture of medicaments for treating infertility, particularly male infertility.

[0176] The present invention also relates to a method for treating infertility, particularly male infertility, comprising contacting sperm cells with a composition or a gel as defined above.

[0177] The present invention also relates to artificial insemination instruments for artificial insemination (AI), in vitro fertilization (IVF) and / or intracytoplasmic sperm injection (ICSI), comprising compositions as defined above or gels as defined above.

[0178] The present invention also relates to the use of compositions or gels as defined above in artificial insemination (AI), in vitro fertilization (IVF), and / or intracytoplasmic sperm injection (ICSI) methods.

[0179] The present invention also relates to a method for artificial insemination of humans and / or animals, including the use of a composition or gel as defined above.

[0180] As defined above, “method” and “use” can be understood as therapeutic or non-therapeutic methods and therapeutic or non-therapeutic uses, in vitro or in vivo methods, and in vitro or in vivo uses. Attached Figure Description

[0181] Figure 1 The graphs show the sperm cell concentration, sperm cell motility, and active sperm cell concentration in the test samples at T=2H and T=4H.

[0182] Figure 2 This is a graph showing the sperm cell viability in the test samples at T=0 and T3H 37℃.

[0183] Figure 3 The graph shows the sperm cell viability in the test samples at T=0 and T3H 37℃.

[0184] Example

[0185] Example 1 (Anti-settling effect):

[0186] Materials and methods:

[0187] Objective: To evaluate sperm cell release at 37°C using the compositions of the present invention.

[0188] - 4 bull semen samples (from the CIA, Coopérative Interdépartementale) et d'InséminationAnimale, L'Aigle) were mixed with OptiXcell Alpha at 4°C and diluted to approximately 200 × 10⁻⁶. 6 sperm / mL.

[0189] - A temperature-responsive hydrogel made of HA-g-PNiPAm (concentration 2%) dissolved in OptiXcellAlpha at 4°C.

[0190] - The sample was homogenized at 4°C.

[0191] - The mini straws were filled and frozen.

[0192] Pour the contents of the well plate into the 6 pipettes and store at 37°C.

[0193] Add 1 mL of EHS (embryo preservation solution) to each well.

[0194] -Incubate the pores at 37°C.

[0195] - To assess sperm release, the supernatant from the well was aspirated using a pipette and analyzed.

[0196] - Use IVOS II at T=2H and T=4H TM Sperm motility analysis was performed using CASA (Computer-Assisted Sperm Analysis) technology.

[0197] The control group corresponds to using only OptiXcellAlpha.

[0198] Results:

[0199] After clearing the wells: the gel can be clearly observed.

[0200] The results are as follows Figure 1 As shown.

[0201] like Figure 1 As shown, the HA-g-PNiPAm gel exhibited interesting viability results during pipetting of the supernatant in the tube, indicating that CASA only analyzed motile sperm cells. In contrast, the control group (using OptiXcellAlpha only) showed 35% motile sperm cells after 2 hours at 37°C. A similar trend was observed regarding motile sperm concentration: HA-g-PNiPAm showed higher values, approximately 95 million sperm after 4 hours at 37°C, compared to 61 million sperm / mL in the control group.

[0202] HA-g-PNiPAm showed the release of active sperm cells over time.

[0203] Conclusion:

[0204] The controlled release of sperm cells was successfully achieved within 4 hours.

[0205] Example 2 (Photocleavage chelator):

[0206] The inventors have discovered that DMNP-EDTA is a photolytic chelating agent that can release divalent ions after light exposure (external stimulation), thereby achieving cross-linking of alginate and capturing sperm.

[0207] Materials and methods:

[0208] -DMNP-EDTA (PromoKiné)

[0209] -BaCl2(Sigma)

[0210] -Algaia alginate (M / G ratio approximately 0.7-0.8),

[0211] -water

[0212] A 1.2 mM BaCl2 solution was prepared in OptiXcell.

[0213] - Resuspend 1 mL of this solution in a 5 mg DMNP-EDTA vial.

[0214] - Disperse 0.125g of alginate in 5mL of BaCl2 / DMNP-EDTA / water solution.

[0215] - The gel was cast into a spectrophotometer and then irradiated with a 395nm ultraviolet lamp for 1 minute.

[0216] The control group corresponds to the solution that was not exposed to light.

[0217] Results:

[0218] The gel is neither too viscous to be cast nor too thin, and it has a good visual gelling effect.

[0219] After being exposed to light, Ba 2+ Ions are released, forming alginate cross-links.

[0220] In contrast, the control group (the unexposed solution) sank and remained in a viscous liquid state, while the gel treated with flash was firmer and more elastic.

[0221] Toxicity test - Materials and methods:

[0222] - Prepare a 2.4 mM BaCl2 solution in OptiXcell.

[0223] - Resuspend 1 mL of this solution in a 5 mg DMNP-EDTA vial.

[0224] Add 1.5 mL of DMNP-EDTA to bring the final concentration of DMNP-EDTA to 2.6 mM.

[0225] Store the solution away from light.

[0226] -Contact the solution with semen at different chelating agent / semen ratios:

[0227] 1:1 ratio => 500 μL chelating agent + 500 μL bull semen (concentration of 57 million sperm / mL)

[0228] 1:0.75 => 500μL chelating agent + 375μL bull semen (concentration of 57 million sperm / mL)

[0229] 1:0.5 => 500μL chelating agent + 250μL bull semen (concentration of 57 million sperm / mL)

[0230] 1:0.25 => 500μL chelating agent + 125μL bull semen (concentration of 57 million sperm / mL)

[0231] -Preparation of a solution using only BaCl2: 500 μL 2.4 mM BaCl2 + 500 μL bull semen with 57 million sperm / mL

[0232] -The following analysis was performed on frozen bull semen:

[0233] - Using IVOS II at T=0 and T3H 37℃ TM Perform vitality analysis.

[0234] - Acrosome integrity analysis was performed on EasyCyte at T=0 and T3H 37℃.

[0235] - Survival analysis of EasyCyte at T=0 and T3H 37℃

[0236] Mitochondrial activity was analyzed on EasyCyte at T=0 and T3H 37℃.

[0237] The control group corresponds to using only OptiXcell.

[0238] Results - Viability and survival rate:

[0239] The results are as follows Figure 2 and Figure 3 As shown.

[0240] like Figure 2 As shown, the presence of the chelating agent did not affect sperm motility. At T=3H, the 1 / 1 (chelating agent / sperm) condition was superior to the control group (9 percentage points higher) (using OptiXcell only).

[0241] like Figure 3 As shown, at T=0, the presence of the chelating agent had a positive effect on sperm viability. At T=0, the 1 / 0.75 condition proved to be optimal in terms of viability (13 percentage points higher than the control group). At T=3H, a ratio range effect began to appear, and viability was affected as the chelating agent / sperm ratio decreased. However, the BaCl2 condition remained unchanged.

[0242] Conclusion:

[0243] The presence of the chelating agent at 37°C (T0 and T3H) did not have a significant impact on viability and survival rate.

Claims

1. A composition comprising: - Human and / or animal semen contains sperm cells, such as sperm. -A compound selected from collagen peptides, polymers and copolymers including polysaccharides, and mixtures thereof. - Semen diluent -Optionally, Slo3 potassium channel inhibitors, and -Optionally, lipids, such as liposomes.

2. The composition of claim 1, wherein the polymer and copolymer are selected from: anionic polymers and copolymers, including anionic polysaccharides; cationic polymers and copolymers, including cationic polysaccharides; nonionic polymers and copolymers, including nonionic polysaccharides; covalently grafted polymers and copolymers, such as polymers and copolymers grafted with amino functional groups; thermally responsive or temperature-responsive polymers and copolymers, such as LCST polymers and copolymers and UCST polymers and copolymers, including polymers and copolymers grafted from polysaccharides, grafted onto polysaccharides, or grafted through polysaccharides; and mixtures thereof.

3. The composition according to claim 1 or 2, wherein the polymer and copolymer are selected from the following substances: polysaccharides, gelatin, ethylene glycol methacrylate polymer (PEGMA), triethylene glycol methacrylate polymer (PTEGMA), polyoligo(ethylene glycol) methyl ether methacrylate polymer (POEGMA), N-isopropylacrylamide polymer (PNiPAM), poly-N-vinylcaprolactam polymer (PNVCL), poloxamer, etc. F-127 (CAS 9003-11-6), poly(ethylene oxide-co-propylene oxide) polymer (PEPO), covalently grafted polymers and copolymers such as polymers and copolymers grafted with amino functional groups, including amino-terminated poloxamer, amino-terminated poly(N-isopropylacrylamide) (PNiPAM), such as amino-terminated poly(N-isopropylacrylamide) (PNiPAM) using cysteine, amino-terminated poly(ethylene oxide-co-propylene oxide) (PEPO), amino-terminated poly(N-vinylcaprolactam) (PNVCL), amino-terminated ethylene glycol methacrylate polymer (PEGMA), amino-terminated polyoligo(ethylene glycol) methyl ether methacrylate polymer (POEGMA), amino-terminated polyoxazoline, and mixtures thereof.

4. The composition according to any one of claims 1 to 3, wherein the polysaccharide is selected from: gums, such as guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum, gellan gum, and mixtures thereof; cellulose, such as microcrystalline cellulose (MCC) and cellulose derivatives, such as methylcellulose, carboxymethyl cellulose (CMC), hydroxymethyl cellulose, ethylcellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose (HPMC), and mixtures thereof; alginate, such as sodium alginate; pectin; pullulan; chitosan; hyaluronic acid (HA); dextran; sclerotium polysaccharide; and mixtures thereof.

5. The composition according to any one of claims 1 to 4, wherein the composition further comprises metal ions, preferably divalent and / or trivalent metal ions, more preferably selected from beryllium ions, magnesium ions, calcium ions, strontium ions, barium ions, radium ions and mixtures thereof.

6. The composition according to any one of claims 1 to 5, wherein the concentration of said compound is from 0.001% to 10% by weight relative to the total weight of said composition, preferably from 0.01% to 10% by weight, more preferably from 0.1% to 5% by weight.

7. The composition according to any one of claims 1 to 5, wherein the concentration of said compound is from 0.001% to 5% by weight relative to the total weight of said composition, preferably from 0.01% to 4% by weight, more preferably from 0.1% to 3% by weight.

8. The composition according to claim 7, wherein the compound is selected from: gelatin, microcrystalline cellulose (MCC), methylcellulose, carboxymethyl cellulose (CMC), hydroxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose (HPMC), pullulan, dextran, gums such as guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum and mixtures thereof, sclerotium polysaccharide, peptan, and mixtures thereof.

9. The composition according to any one of claims 1 to 5, wherein the concentration of said compound is 0.1% to 10% by weight of the total weight of the composition, preferably 0.5% to 5% by weight, more preferably 1% to 3% by weight.

10. The composition according to claim 9, wherein the compound is selected from: methylcellulose, carboxymethylcellulose (CMC), hydroxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose (HPMC), alginate, pectin, pullulan, chitosan, hyaluronic acid (HA), dextran, gums such as guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum and mixtures thereof, peptan, polymers and copolymers such as ethylene glycol methacrylate polymer (PEGMA), triethylene glycol methacrylate polymer (PTEGMA), polyoligo(ethylene glycol) methyl ether methacrylate (POEGMA), N-isopropylacrylamide polymer (PNiPAM), poly-N-vinylcaprolactam (PNVCL), poloxamer, etc. F-127 (CAS 9003-11-6), poly(ethylene oxide-co-propylene oxide) polymer (PEPO), and mixtures thereof.

11. The composition according to claim 9, wherein the compound is selected from: carboxymethyl cellulose (CMC), alginate, pectin, chitosan, hyaluronic acid (HA), gums such as guar gum, xanthan gum, gum arabic, carrageenan, arabinogalactan gum, and mixtures thereof, and is combined with a metal ion, wherein the metal ion is preferably selected from beryllium ions, magnesium ions, calcium ions, strontium ions, barium ions, radium ions, and mixtures thereof.

12. The composition according to any one of claims 1 to 11, wherein the composition further comprises a divalent or trivalent ion chelating agent, preferably a photolytic divalent or trivalent ion chelating agent, more preferably selected from o-nitrobenzyl compounds, such as o-nitrobenzyl-BAPTA and DMNP-EDTA, and mixtures thereof.

13. The composition according to claim 12, wherein the concentration of the chelating agent ranges from 0.005% to 5% relative to the total weight of the composition, preferably from 0.01% to 3% relative to the total weight of the composition, and more preferably from 0.05% to 1% relative to the total weight of the composition.

14. A method for manufacturing the composition according to any one of claims 1 to 13, comprising the following steps: - The semen diluent is mixed with a compound, optionally a metal ion, an Slo3 potassium channel inhibitor, and / or a lipid, such as a liposome, and optionally additional compounds and / or divalent or trivalent ion chelating agents to obtain a mixture. - Dilute the mixture with water to obtain a diluted mixture, and - Mix human and / or animal semen into the diluted mixture.

15. Use of the composition according to any one of claims 1 to 13 in increasing the contact time between sperm cells and Slo3 potassium channel inhibitors.

16. Use of the composition according to any one of claims 1 to 13 in preventing sperm cell precipitation.

17. The composition according to any one of claims 1 to 13, for improving the motility, survival rate and lifespan of sperm cells, particularly sperm, preferably capacitated sperm.

18. The composition according to any one of claims 1 to 13, for improving fertility.

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