TECHNICAL FIELD

The invention falls within the technical fields of molecular neuropharmacology and relates to a biocompatible Humanin-loaded solid lipid nanoparticle which allows the active ingredient to cross the blood-brain barrier in order to provide treatment to patients suffering from neurological diseases and increases the effectiveness of the treatment by enabling more active ingredient to pass through the disease site, to the production thereof and to the use thereof in the treatment of neurology diseases or in the symptoms arising from these diseases.

In another aspect, the invention provides a method for the production of Humanin- loaded solid lipid nanoparticles.

PRIOR ART

Molecular neuropharmacology is a science investigating the neural mechanisms that affect the cellular function and behavior of drugs in the nervous system. The general object of Molecular neuropharmacology is to study neurons and neurochemical interactions, as well as developing drugs that have beneficial effects on neurological function. As it is better known, this field relates to the investigation of the interactions of neurotransmitters, neuropeptides, neurohormones, neuromodulators, enzymes, second messengers, co-transporters, ion channels and receptor proteins in the central and peripheral nervous systems. Some of the diseases studied by neuropharmacology are epilepsy, Alzheimer's Disease, Dementia, Migraine-headaches, Parkinson's Disease and sleep disorders of various severities.

According to the World Alzheimer Report published in 2015, 46.5 million people in the world have dementia, and this number doubles every 20 years. This number is estimated to exceed 100 million by 2050. The medical condition that develops as memory loss, second childhood (Dementia) and in general terms, a decrease in cognitive functions due to the death of brain cells over time is called Alzheimer's disease. Alzheimer's Disease, a neurological disease, is also the most common type of dementia. Beta amyloid plaques are seen in the brain of people with the disease. The disease, which manifests itself only with simple forgetfulness in the initial phase, can progress over time to the point where the patient forgets recent events and cannot recognize family members and immediate circle. During further advanced stages of the disease, patients have difficulty meeting their basic needs and become in need of care. Alzheimer's patients usually apply to clinics with complaints of decreased performance in cognitive and behavioral areas. Although the symptoms in the initial stage of the disease are milder, the findings are more pronounced in patients in the advanced stage. Although the symptoms of Alzheimer's onset are generally minor memory problems, they include symptoms such as forgetting recent conversations and events, and not being able to remember the names of people, objects and places. The most common Alzheimer's symptoms as the disease progresses are: confusion, difficulty in adapting to one's environment, getting lost in places where one knows well, problems with speech and language skills, aggression, making unusual demands from family and friends, the appearance of personalisty disorders such as suspicion of the environment, hallucinations and delusions, low motivation and self-esteem, difficulty in performing daily activities without help, denial of events that the person cannot remember, anxiety and depression. The symptoms mentioned are the symptoms which are often seen when the disease is first diagnosed. As the disease progresses, these symptoms increase in severity and reach much more advanced levels, such as the patient not being able to recognize family members, completely forgetting his/her recent past, and having difficulty even recognizing himself/herself. In this case, patients often need a caregiver to continue their daily lives.

As is known in the art, drug forms containing the active ingredients donepezil, rivastigmine, galantamine, which are an acetylcholinesterase inhibitor, and memantine which is an N-Methyl-D-aspartate receptor antagonist (NMDA), are currently used in the treatment of Alzheimer's disease, however, the drugs used cannot change the developing symptoms and prevent the progression of the disease. The formation of amyloid plaques, which is seen in Alzheimer's disease and is discussed as the cause of the disease, is now considered as a starting point from which the studies may be carried out to prevent the progression of the disease. The biggest blockage to achieve a success in the treatment of neural diseases is the Blood Brain Barrier (BBB for short) in the brain. The existence of a selectively permeable barrier between blood and brain was first detected by Paul-Ehrlich in 1880. BBB is a series of semi-permeable specialized endothelial cell membranes which allow the passage of necessary nutrients while protecting the brain from harmful substances and toxins circulating in the blood to ensure hemostasis in the brain. BBB prevents the passage of compounds from the blood to the extracellular environment of the brain. While 98% of small molecule active ingredients cannot cross the BBB, almost all large molecule active ingredients cannot cross the BBB. In order for an active ingredient to cross the BBB, it should be lipid soluble, be non-ionized at a physiological pH, have a low molecular weight, and have low binding rates to serum proteins.

In the relevant technical field, studies are ongoing on the investigation of treatment methods which can treat neural diseases such as Alzheimer's Disease with high efficiency and enable the passage of high amounts of active ingredients by crossing the Blood-Brain Barrier which poses a major technical problem during treatment, and it has become a necessity to provide innovations in the mentioned technical field.

BRIEF DESCRIPTION OF THE INVENTION

The invention relates to an active ingredient-loaded solid lipid nanoparticle for the treatment of neural diseases such as Alzheimer's Disease and Dementia and the relief of symptoms arising from these diseases.

As is known in the art, technical problems encountered in the treatment of neural disorders such as Alzheimer's Disease and Dementia are that the drugs used in treatments cannot change the developing symptoms and prevent the progression of the disease. One of the biggest problems encountered in the treatment of neural disorders is that the BBB cannot be crossed at a high rate in people with disease, and the therapeutic active ingredient cannot interact sufficiently with the disease sites. The main object of the invention is to ensure the high amounts of active ingredient to interact with the diseased site by exceeding the BBB in patients who pose a technical problem during the treatment. In addition, the invention includes research and development activities to configure the active ingredient-loaded solid lipids to have a high interaction with disease sites. Another object of the invention is to provide a method for the production of configured active ingredient-loaded solid lipid nanoparticles which may be used in the treatment of neural disorders such as Alzheimer's Disease and Dementia. As mentioned before, the inventors have provided configurations so that the active ingredient-loaded solid lipid nanoparticle of the invention may have a high interaction with the disease site after crossing the BBB with high amounts.

Another object of the invention is to use the obtained active ingredient-loaded solid lipid nanoparticles in neural diseases and in relieving the symptoms arising from these diseases.

In order to achieve all these, the present invention relates to a Humanin-loaded solid lipid nanoparticle which is suitable for use in the treatment of neurological diseases such as Alzheimer's Disease and Dementia or in relieving the symptoms arising from these diseases and allows the blood-brain barrier to be crossed with high efficiency, thus allowing a high amount of therapeutic active ingredient to interact with the diseased site, characterized in that it comprises at least one Humanin-based therapeutic molecule as the active ingredient and a solid lipid nanoparticle loaded with said active ingredient.

A possible embodiment of the invention is that the Humanin-based molecule is Humanin and/or Gly14-Humanin.

A possible embodiment of the invention is that the Humanin-based molecule is Gly14 Humanin.

A possible embodiment of the invention is that it is modified with at least one of transferrin, lactoferrin, folic acid, 83-14 monoclonal antibodies, betahydroxybutyric acid and apolipoprotein E ligands.

A possible embodiment of the invention is that it is modified with the transferrin ligand.

A possible embodiment of the invention is that it comprises at least one auxiliary active ingredient. In another aspect, the invention relates to a pharmaceutical composition comprising a Humanin-loaded solid lipid nanoparticle as a component, wherein the pharmaceutical composition is suitable for use in the treatment of neurological diseases such as Alzheimer's Disease and Dementia or in relieving the symptoms arising from these diseases and allows the blood-brain barrier to be crossed with high efficiency, thus allowing a high amount of therapeutic active ingredient to interact with the diseased site.

In another aspect, the invention relates to a method of producing solid lipid nanoparticles loaded with the Gly14-Humanin and/or Humanin, the active ingredients, wherein the method comprises the following process steps:

- heating the lipid(s) to a temperature in the range of 70 °C to 80 °C,

- also heating a surfactant and an oil phase to a temperature in the range of 70 °C to 80 °C and adding the lipid(s) at the specified temperature to the oilsurfactant at the specified temperature,

- adding Gly14-Humanin, the active ingredient, to the oil phase after the oil and surfactant are completely melted and mixing in a magnetic stirrer for at least 20 seconds to thereby ensure a homogeneous dispersion in the oilsurfactant phase,

- also heating the water phase to a temperature in the range of 70 °C to 80 °C,

- adding the water phase at the specified temperature to the oil-lipid- surfactant phase at the specified temperature and mixing with a mixer until homogenization is achieved and obtaining an emulsion,

- sonicating the resulting emulsion and performing cooling processes.

A possible embodiment of the invention is that said lipid(s) is/are at least one selected from the group consisting of glyceryl monostearate, glyceryl distearate, glyceryl tristearate, myristin, monopalmitin, tripalmitin, glyceryl behenate, trilaurin, and cetyl palmitate. A possible embodiment of the invention is that said surfactant is at least one selected from the group consisting of poloxamer, polysorbate, polyoxyethylene aliphatic alcohol ether, and polyoxyethylene fatty acid ester group.

A possible embodiment of the invention is that the oil phase is Tween 80.

A possible embodiment of the invention is that the amount of lipid is added to the oil phase at a ratio of 2/3 by weight (w/w) as lipid/oil .

In another aspect, the invention relates to a method of producing solid lipid nanoparticles loaded with the Tf-modified Gly14-Humanin and/or Humanin, the active ingredients, wherein the method comprises the following process steps:

- adding DSPE-PEG(2000) amine to the lipid(s) phase,

- heating DSPE-PEG(2000) amine-lipid(s) to a temperature in the range of 70 °C to 80 °C,

- also heating a surfactant and an oil phase to a temperature in the range of 70 °C to 80 °C and adding the lipid(s) at the specified temperature to the oilsurfactant at the specified temperature,

- adding Gly14-Humanin, the active ingredient, to the oil phase after the oil and surfactant are completely melted and mixing in a magnetic stirrer for at least 20 seconds to thereby ensure a homogeneous dispersion in the oilsurfactant phase,

- also heating the water phase to a temperature in the range of 70 °C to 80 °C,

- adding the water phase at the specified temperature to the oil-lipid- surfactant phase at the specified temperature and mixing with a mixer until homogenization is achieved and obtaining an emulsion,

- sonicating the resulting emulsion and performing cooling processes,

- treating 1 -ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) with a binding agent to activate the end thereof to which transferrin will be bound, wherein the treatment process mentioned herein comprises mixing Tf with EDC at room temperature for at least 20 minutes, wherein the amount of Tf is in the range of 18/1 to 19/1 by weight (w/w) as lipid/Tf, and the amount of EDC is between 2/1 (ml/mg) to 1/1 (ml/mg) on a volume/weight basis,

Obtaining a suspension of Tf and EDC mixture in a buffer solution with the active ingredient-loaded SLN formulation comprising DSPE-PEG(2000)- amine.

A possible embodiment of the invention is that said buffer solution is a phosphate- based buffer solution with a pH value of 7.4.

A possible embodiment of the invention is that said suspension process is carried out with mixing at room temperature for at least 2 hours.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1-A shows a graph of determining the particle sizes of the solid lipid nanoparticles.

Figure 1-B shows a graph for the surface load of the solid lipid nanoparticles.

Figure 2-A shows a TEM image of the Humanin-loaded solid lipid nanoparticles.

Figure 2-B shows a TEM image of the Tf-modified Humanin-loaded solid lipid nanoparticles.

DETAILED DESCRIPTION OF THE INVENTION

In this detailed description, the invention falls within the technical fields of molecular neuropharmacology; relates to a biocompatible Humanin-loaded solid lipid nanoparticle which allows the active ingredient to cross the blood-brain barrier in order to provide treatment to patients suffering from neurological diseases and increases the effectiveness of the treatment by enabling more active ingredient to pass through the disease site, to the production thereof and to the use thereof in the treatment of neurology diseases or in the symptoms arising from these diseases; and is described by the examples for a better understanding of the subject without any limiting effect. In the art, a treatment approach for neural diseases such as Alzheimer's Disease and Dementia is to use Humanin molecules as active ingredients of a peptide structure, which have recently been proven to inhibit the formation of amyloid plaques and thus make possible a radical treatment of neural diseases such as Alzheimer's Disease and Dementia. Another name for Ap plaques is senile plaques and they consist of fibril forms of Ap peptides. Senile plaques are structures formed from amyloid precursor protein (APP) proteolytically, a large transmembrane protein which is 40-42 amino acids in length and accumulates outside the cell, whose function is not fully understood. Ap plaques seen in the brains of Alzheimer's patients mostly contain Ap(1 - 40) and AP(1 -42). It means that the C terminus ends at amino acids 40 and 42. The one that causes more plaque formation is the 42-amino acid form as it is the first version to precipitate. Amyloid p then aggregates into diffusible plaques, which turn into dense senile plaques. These plaques cause inflammation, synaptic loss, abnormal phosphorylation, neurofibrillary tangle formation, and cell death. This condition is a pathological finding of clinical dementia.

The molecule referred to as "Humanin" in the invention is a micropeptide encoded in the mitochondrial genome by the 16S ribosomal RNA gene, MT-RNR2. Its structure contains a three-turn a helix and no symmetry. The sequence of Humanin is H ₂ N-Met- Ala-Pro-Arg-Gly-Phe-Ser-Cys-Leu-Leu-Leu-Leu-Thr-Ser-Glu-lle- Asp-Leu-Pro-Val-Lys- Arg-Arg-Ala-OH and has a chemical formula of C119H204N34O32S2 (referred as Formula 1 ). Humanin has been detected both in the circulation and in cell membranes. It has been found in many cell types under physiological conditions, including endothelial cells, glial cells, and germ cells. It shows its effects through specific receptors. It has a high metabolic rate and is localized in many tissues, and its expression decreases with age. When humanin peptide is transfected into a neuron cell, it protects the cell against cytotoxicity caused by amyloid beta (AP)-42. It is known in the art that both synthetic and secreted Humanin exhibit this effect and that the Humanin effect also includes cellular receptors.

Formula 1 .

In a preferred embodiment of the invention, the active ingredient-loaded solid lipid nanoparticle comprises Gly14-Humanin as the active ingredient. Gly14-Humanin is a variant of Humanin and is obtained by substitution of Glycine for Ser14 and has been shown to be 1000 times more active than Humanin. The sequence of the Gly14- Humanin peptide is as follows: H-Met-Ala-Pro-Arg-Gly-Phe-Ser-Cys-Leu-Leu-Leu-Leu- Thr-Gly-Glu-lle-Asp-Leu-Pro-Val-Lys-Arg-Arg-Ala-OH.

In the invention, Gly14-Humanin and/or Humanin, as the active ingredients, or the pharmaceutically acceptable salts thereof should be delivered to the disease site in order to treat neural diseases such as Alzheimer's Disease and Dementia and to relieve the symptoms caused by these diseases.

In the art, the biggest problem for the pharmaceutical active ingredients to reach the site of neural disease is to ensure the BBB part is crossed with high efficiency. Accordingly, the present inventors have determined that the drug delivery system to which the active ingredients will be loaded should be compatible with the BBB part and relatively small in size. The present inventors have determined that the drug delivery system should be compatible with the BBB. Accordingly the drug delivery system is expected to be lipid soluble, be non-ionized at physiological pH values, have a low molecular weight, and have low binding to serum proteins. The present inventors propose the use of the solid lipid nanoparticles as a drug delivery system. In the invention, the solid lipid nanoparticles (known to be abbreviated as SLN) are referred to as new nanoparticulate drug delivery systems prepared from lipids that are solid at body temperature. SLNs are aqueous colloidal dispersions obtained with biocompatible and biodegradable lipids formed into a compact matrix to which various active ingredients may be loaded. SLNs are similar to the lipophilic BBB in terms of their structure, and their relatively smaller size provides an advantage in the transport of active ingredients to the brain and the crossing of the BBB. Many studies have shown that they are suitable transporters for peptide protein substances and that these substances increase the stability and the active ingredient release performance. In the invention, the present inventors do not determine the scope of protection regarding the production or obtaining of the SLNs, the SLNs suitable for use in the relevant technical field fall within the scope of protection of the present invention.

The innovative aspect of the present invention is that Humanin or Gly14-Humanins, or the mixtures thereof in certain weight ratios are loaded to the solid lipid nanoparticles which are compatible with the BBB part and have a high permeability therethrough in order for them to interact with the disease sites with high efficiency for the treatment of Dementia and Alzheimer's diseases or for relief of the symptoms arising from these diseases. In this way, the main embodiment of the present invention is the solid lipid nanoparticles loaded with the active ingredients, Gly14-Humanin and/or Humanin, for the treatment of Dementia and Alzheimer's diseases or the relief of symptoms arising from these diseases.

A preferred embodiment of the present invention is to modify the solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, with the agents suitable for the brain structure to increase the interaction with the disease site. Modification of the solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, is to provide the chemical configuration of at least the surface part of the molecule by adding agents to its structure and to increase the affinity in accordance with the chemical in the disease site. At least one of the ligands transferrin (abbreviated as Tf), lactoferrin (abbreviated as Lf), folic acid, 83- 14 monoclonal antibodies, betahydroxybutyric acid and apolipoprotein E is included in the invention as said agent. In a preferred embodiment of the present invention, the solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, are modified with an agent, wherein said agent is a Tf ligand.

In the invention, the solid lipid nanoparticles loaded with the active ingredient(s), Gly14- Humanin and/or Humanin, are modified with Tf and targeted to the brain, making it possible to deliver more active ingredient to the disease site. Transferrin receptors are among the most highly expressed receptors in the blood-brain barrier cell membrane. Iron delivery to the brain is achieved through binding of the iron-binding protein, transferrin (Tf), and uptake into the cell by the receptor-mediated transcytosis. Tf contained in the formulation increases drug biodistribution into the brain by increasing drug uptake across the person's blood-brain barrier through the Tf receptor (TfR).

Accordingly, the main innovative aspect of the present invention is related to obtaining the solid lipid nanoparticles modified with the Tf ligand loaded with the active ingredient(s), Gly14-Humanin and/or Humanin.

The invention offers a method for the production of the solid lipid nanoparticles loaded with the active ingredients, Gly14-Humanin and/or Humanin. Accordingly, the method of the invention includes the following process steps:

§ heating the lipid(s) to a temperature in the range of 70 °C to 80 °C,

§ also heating a surfactant to a temperature in the range of 70 °C to 80 °C and adding the lipid(s) at the specified temperature to the oil at the specified temperature,

§ adding Gly14-Humanin, the active ingredient, to the oil phase after the oil and surfactant are completely melted and mixing in a magnetic stirrer for at least 20 seconds to thereby ensure a homogeneous dispersion in the oil phase,

§ also heating the water phase to a temperature in the range of 70 °C to 80 °C,

§ adding the water phase at the specified temperature to the oil-lipid phase at the specified temperature and mixing with a mixer until homogenization is achieved and obtaining an emulsion,

§ sonicating the resulting emulsion and performing cooling processes. Said lipid(s) is/are at least one selected from the group consisting of glyceryl monostearate, glyceryl distearate, glyceryl tristearate, myristin, monopalmitin, tripalmitin, glyceryl behenate, trilaurin, and cetyl palmitate.

Said emulsifier and surfactant are at least one selected from the group consisting of poloxamer, polysorbate, polyoxyethylene aliphatic alcohol ether, and polyoxyethylene fatty acid ester group.

Tween 80 oil is used as the oil mentioned in the invention.

The lipid(s) mentioned here are added to the oil at a ratio of 2/3 by weight as lipid/oil in order to form an oil phase.

In the invention, the mixing process for the homogenization process is carried out at a speed between 10000 to 50000 rpm. In a preferred embodiment, this value is between 15000 rpm to 25000 rpm.

In the invention, ultrasonic probes for sonicating the emulsion are expected to have a strength in the range of 80% to 95%. Said sonication process is carried out for a period of time between 1 to 20 minutes. In a preferred embodiment, the sonication process is carried out for a period of time between 5 to 15 minutes. In this way, nanoparticles are obtained in an appropriate size range.

In a preferred embodiment of the invention, the particles obtained after sonication and cooling are processed in a laminar air flow cabin. In this way, the particles are allowed to be sterile.

In the most preferred embodiment of the invention, the solid lipid nanoparticles loaded with the active ingredients, Gly14-Humanin and/or Humanin, are modified with Tf agents. In another aspect, the invention relates to a method of modifiying the solid lipid nanoparticles loaded with the active ingredients, Gly14-Humanin and/or Humanin, with Tf agents. In the invention, SLNs are reproduced using DSPE-PEG(2000) amine (1 ,2- distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyeth ylene glycol)-2000] (ammonium salt) for Tf binding to SLNs. The modified method of the invention includes the following process steps: § adding DSPE-PEG(2000) amine to the lipid phase of the SLN,

§ treating 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) with a binding agent to activate the end thereof to which transferrin will be bound, wherein the treatment process mentioned herein comprises mixing Tf with EDC at room temperature for at least 20 minutes, wherein the amount of Tf is in the range of 18/1 to 19/1 by weight as lipid/Tf, and the amount of EDC is between 2/1 (ml/mg) to 1/1 (ml/mg) on a volume/weight basis,

§ Obtaining a suspension of Tf and EDC mixture in a buffer solution with the active ingredient-loaded SLN formulation comprising DSPE-PEG(2000)-amine.

Said buffer solution is 0.25 mL of phosphate solution with pH 7.4. For the suspension process, it is mixed at room temperature for at least 2 hours.

In addition, in the invention, the uncombined Tf excess during these processes is removed by a dialysis membrane method.

The size and surface charge analysis measurements are performed for the Tf ligand- modified solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, and the solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, which are obtained by applying the process steps of the method mentioned in the invention (including the modifying steps of the method). In the invention, the particle size (hydrodynamic diameter) and the surface charge were analyzed by Dynamic Light Scattering (DLS) and electrophoretic light scattering methods, respectively, using a Zetasizer Nano ZS device. Data regarding the obtained nanoparticle size are shown in Figure 1 -A and the data regarding the surface charge are shown in Figure 1-B.

The average hydrodynamic diameter of the humanin-loaded SLNs was determined as 104.90 ± 0.92 nm and the PDI distribution was determined as 0.29 ± 0.01. For particles modified with Tf, these values were found to be 117.40 ± 1.16 nm and 0.39 ± 0.03, respectively. The increase in particle size is considered normal due to the binding of a large group such as Tf to a particle. Zeta potentials were found to be -15.0±096 for Hum-SLN and -9.10±0.39 for Tf-Hum-SLN. The bound Tf has been determined to reduce the zeta value.

The encapsulation efficiency was obtained as 98.99% for Humanin-loaded SLN and 99.97% for Tf-modified Humanin-loaded SLN. The rates of substances loaded into the SLN were quite high, and the modification did not cause any change in loading efficiency.

When TEM images of SLNs are examined, the spherical nanoparticles with smooth surfaces are observed to be obtained. In addition, in all formulas, the particle size observed by TEM was found to be similar to the average particle size measured by the photon correlation spectroscopy method, and in this respect, these two data supported each other. Based on these images, the nanoparticles were proven to be successfully prepared.

As the hydrodynamic diameter values of Hum-SLN and Tf-Hum-SLNs are in the range of 100-200 nm, they have been determined to be suitable in terms of dimensions for crossing the BBB.

Figures 2-A and 2-B show the transmission electron microscopy images of the solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, and the Tf ligand-modified solid lipid nanoparticles loaded with the active ingredient(s), Gly14-Humanin and/or Humanin, respectively. Imaging was done by negative staining of the samples using phosphotungstic acid. Briefly, this process consisted of incubating 5 pL of the formulation on carbon-coated grids for 2 minutes, then treating with 0.75% (w/v) phosphotungustic acid staining solution at pH 7.4 for 15 seconds. Analysis was performed after removing excess solution with a filter paper and drying the samples under vacuum.

With the tests carried out within the scope of the invention, the present inventors have determined that the Humanin-SLN and Tf-Humanin-SLN systems can be used effectively for crossing the BBB. Humanin of a peptide structure, which will be transported with the use of said systems, will be able to further cross the BBB and remain stable in the brain environment for a longer time. It is not possible to deliver the substances of a peptide structure directly into the bloodstream due to the enzymes in the blood that break down peptides. Humanin and the similar active materials of a peptide structure, which are encapsulated with the systems developed within the scope of the invention may be transported to a desired location while being protected from the enzymes and the immune system members in the blood circulation. Thus, with further studies, the lipidic system has a potential to be used in treatment. The scope of protection of the invention is specified in the attached claims and cannot be limited to what is explained in this detailed description for the exemplary purposes. That is because it is clear that a person skilled in the art may represent similar embodiments in the light of what has been explained above without departing from the main spirit of the invention.