US20230091957A1 - Method for the Stimulation of Human Immune Cells - Google Patents
Method for the Stimulation of Human Immune Cells Download PDFInfo
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- US20230091957A1 US20230091957A1 US17/993,633 US202217993633A US2023091957A1 US 20230091957 A1 US20230091957 A1 US 20230091957A1 US 202217993633 A US202217993633 A US 202217993633A US 2023091957 A1 US2023091957 A1 US 2023091957A1
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- 238000000034 method Methods 0.000 title claims abstract description 16
- 210000002865 immune cell Anatomy 0.000 title description 3
- 230000000638 stimulation Effects 0.000 title 1
- 239000000126 substance Substances 0.000 claims abstract description 14
- 230000004936 stimulating effect Effects 0.000 claims abstract description 6
- 210000000987 immune system Anatomy 0.000 claims abstract description 5
- 239000006228 supernatant Substances 0.000 claims description 23
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 241000015203 Saccharina angustata Species 0.000 claims description 5
- 241000015177 Saccharina japonica Species 0.000 claims description 5
- 239000008240 homogeneous mixture Substances 0.000 claims description 5
- 235000012054 meals Nutrition 0.000 claims description 5
- 229910052786 argon Inorganic materials 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 241000195493 Cryptophyta Species 0.000 claims description 3
- 238000002386 leaching Methods 0.000 claims description 3
- 239000013049 sediment Substances 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims 2
- 238000010926 purge Methods 0.000 claims 2
- 238000001291 vacuum drying Methods 0.000 claims 2
- 238000010438 heat treatment Methods 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 2
- 208000026278 immune system disease Diseases 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000002560 therapeutic procedure Methods 0.000 abstract description 2
- 239000003814 drug Substances 0.000 description 14
- 229940079593 drug Drugs 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 9
- 210000002249 digestive system Anatomy 0.000 description 7
- 239000013543 active substance Substances 0.000 description 4
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 4
- 210000003719 b-lymphocyte Anatomy 0.000 description 3
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 2
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 2
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 2
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 2
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 2
- 241001466453 Laminaria Species 0.000 description 2
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 2
- 210000001842 enterocyte Anatomy 0.000 description 2
- 230000004727 humoral immunity Effects 0.000 description 2
- 230000002434 immunopotentiative effect Effects 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 208000006820 Arthralgia Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 206010041349 Somnolence Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000004820 blood count Methods 0.000 description 1
- KTUQUZJOVNIKNZ-UHFFFAOYSA-N butan-1-ol;hydrate Chemical compound O.CCCCO KTUQUZJOVNIKNZ-UHFFFAOYSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000009535 clinical urine test Methods 0.000 description 1
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- 239000012634 fragment Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 210000002175 goblet cell Anatomy 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000036651 mood Effects 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 210000003134 paneth cell Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 230000009993 protective function Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000011369 resultant mixture Substances 0.000 description 1
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- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
- 230000002618 waking effect Effects 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
Definitions
- the invention pertains to the domain of medicine, particularly immunology, and can be used for stimulating cells of the human immune system in treating various diseases involving immune system disorders.
- a known immunopotentiation method uses Panagen (No. LSR-004429/08), administered in the form of tablets or intramuscular solution, that stimulates endogenous production of cytokines and hematopoietic factors (patent: RU 2498821 C1, published on 4 Oct. 2012).
- the drug is double-stranded fragmented human genomic DNA with fragments 200 to 6000 base pairs long.
- This method fails to achieve highly efficient immunopotentiation due to the medicine's low bioavailability, and losses during passage through segments of the digestive system if taken orally.
- it may be actively destroyed; also, being a specific nucleotide sequence, it may induce negative responses by interfering with DNA structure in human cells.
- Panagen is used in tableted form, three tablets per day, containing 5 mg of the active substance, for one to fourteen days since the start of administration or until therapeutic benefit is achieved. If used in tableted form, the foreign DNA-based medicine may lose much of its active substance and efficiency.
- Our proposed method aims to create a medication for stimulating human immune cells that is devoid of the known drug's deficiencies, with higher bio-availability and cell penetration capability due to both its active substances' small size (5-10 kD) and smaller losses during passage through the digestive system on account of the chemical structure of its active components that mainly include polysaccharides.
- a human immune cell stimulant by leaching homogenate of Laminaria Japonica and Laminaria Angustata algae with aqueous and isobutanolic solution, with 1 g:2.5 ml:2.5 ml mix ratio, to obtain a homogeneous mixture which is then cured at 60° C. for one hour and incubated for 10-15 hours at room temperature; then supernatant fluid is withdrawn and centrifuged at 3000 rpm and +4° C. for 30 minutes. The centrifuged supernatant is placed into a vacuum evaporator and blown through with argon, then vacuum-dried for one hour at room temperature and then at 60° C. until all liquid fraction evaporates.
- the resultant sediment is diluted with water into 5% solution and then ultracentrifuged at 100,000 rpm for one hour; the resulting supernatant is withdrawn and centrifuged in 10 kD filter tubes for one hour at 5000 rpm, after which the lower percolated fraction is withdrawn and centrifuged in 5 kD filter tubes for one hour at 5000 rpm. Now the upper fraction detained by the filter is withdrawn and diluted in 500 ml of NaCl saline for a final concentration of 10 mg/ml.
- the centrifuged supernatant is ultracentrifuged at 100,000 rpm for one hour.
- a second aspect of the invention comprises the substance produced by the method described above.
- a further aspect of the invention comprises the substance, which is produced by the method described above, for use in a therapeutic method for stimulating cells in the human immune system.
- this substance is administered orally, preferably in 1 ml dosage once daily, for example before meals, for a fortnight.
- Enterocytes and goblet cells found in extrathoracic trachea and pancreatic and parotid ducts as well as segments of the digestive system, are three times more permeable to 5-10 kD molecules than other digestive system cells, including stem and Paneth cells. Consequently, the cell pools partly responsible for tissue immunity manifestations and the digestive system's protective functions and also for the ingress of substances from the digestive system into the bloodstream turn out to be the cell pools most permeable to the algal extract as compared to others. As a result, the active algal extract can enter the bloodstream both paracellularly and transcellularly (through the enterocytes).
- the proposed method's algorithm is as follows:
- the resulting supernatant should be withdrawn and centrifuged again, now in 10 kD filter tubes, for one hour at 5000 rpm on the Eppendorf Centrifuge 5804 R apparatus.
- the lower percolated fraction is then withdrawn and centrifuged in 5 kD filter tubes, for one hour at 5000 rpm on the Eppendorf Centrifuge 5804 R7 apparatus.
- the upper fraction detained by the filter is withdrawn and 500 ml of NaCl saline is added.
- the medication obtained is used as follows: 1 ml of the medication is diluted in 20 ml of water and ingested once a day before a meal for two weeks.
- the medication consists of 5 to 10 kD molecules.
- her patient noted a subjective improvement in her status, with lesser weakness and fatigue; she found it easier to get up in the morning (while her daily regimen remained the same).
- her immune status showed an increase in her natural killer and B lymphocyte pools that doubled and trebled, respectively.
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Alternative & Traditional Medicine (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Provided herein is a substance for use in a therapeutic method for stimulating cells in the human immune system, a method for producing this substance, and a method of use of the substance for stimulating cells of the human immune system in treating various diseases involving immune system disorders.
Description
- The present application is a divisional of U.S. patent application Ser. No. 17/002,169, filed Aug. 25, 2020, the content of which is incorporated by reference herein in its entirety.
- The invention pertains to the domain of medicine, particularly immunology, and can be used for stimulating cells of the human immune system in treating various diseases involving immune system disorders.
- A known immunopotentiation method uses Panagen (No. LSR-004429/08), administered in the form of tablets or intramuscular solution, that stimulates endogenous production of cytokines and hematopoietic factors (patent: RU 2498821 C1, published on 4 Oct. 2012). The drug is double-stranded fragmented human genomic DNA with fragments 200 to 6000 base pairs long. This method fails to achieve highly efficient immunopotentiation due to the medicine's low bioavailability, and losses during passage through segments of the digestive system if taken orally. On the other hand, like any foreign DNA, it may be actively destroyed; also, being a specific nucleotide sequence, it may induce negative responses by interfering with DNA structure in human cells.
- Panagen is used in tableted form, three tablets per day, containing 5 mg of the active substance, for one to fourteen days since the start of administration or until therapeutic benefit is achieved. If used in tableted form, the foreign DNA-based medicine may lose much of its active substance and efficiency.
- Our proposed method aims to create a medication for stimulating human immune cells that is devoid of the known drug's deficiencies, with higher bio-availability and cell penetration capability due to both its active substances' small size (5-10 kD) and smaller losses during passage through the digestive system on account of the chemical structure of its active components that mainly include polysaccharides.
- To work around that problem, we obtain a human immune cell stimulant by leaching homogenate of Laminaria Japonica and Laminaria Angustata algae with aqueous and isobutanolic solution, with 1 g:2.5 ml:2.5 ml mix ratio, to obtain a homogeneous mixture which is then cured at 60° C. for one hour and incubated for 10-15 hours at room temperature; then supernatant fluid is withdrawn and centrifuged at 3000 rpm and +4° C. for 30 minutes. The centrifuged supernatant is placed into a vacuum evaporator and blown through with argon, then vacuum-dried for one hour at room temperature and then at 60° C. until all liquid fraction evaporates. The resultant sediment is diluted with water into 5% solution and then ultracentrifuged at 100,000 rpm for one hour; the resulting supernatant is withdrawn and centrifuged in 10 kD filter tubes for one hour at 5000 rpm, after which the lower percolated fraction is withdrawn and centrifuged in 5 kD filter tubes for one hour at 5000 rpm. Now the upper fraction detained by the filter is withdrawn and diluted in 500 ml of NaCl saline for a final concentration of 10 mg/ml.
- In a further embodiment, after centrifugation at 3000 rpm and +4° C. for 30 minutes and before the centrifuged supernatant is placed into a vacuum evaporator and purged with argon, the centrifuged supernatant is ultracentrifuged at 100,000 rpm for one hour.
- A second aspect of the invention comprises the substance produced by the method described above.
- A further aspect of the invention comprises the substance, which is produced by the method described above, for use in a therapeutic method for stimulating cells in the human immune system. In a preferred embodiment, this substance is administered orally, preferably in 1 ml dosage once daily, for example before meals, for a fortnight.
- Enterocytes and goblet cells, found in extrathoracic trachea and pancreatic and parotid ducts as well as segments of the digestive system, are three times more permeable to 5-10 kD molecules than other digestive system cells, including stem and Paneth cells. Consequently, the cell pools partly responsible for tissue immunity manifestations and the digestive system's protective functions and also for the ingress of substances from the digestive system into the bloodstream turn out to be the cell pools most permeable to the algal extract as compared to others. As a result, the active algal extract can enter the bloodstream both paracellularly and transcellularly (through the enterocytes). The greater permeability of certain digestive system cells and the algal extract's small particle size (5-10 kD) and lack of a complicated 3D structure combine to make it highly bioavailable. Studies of the medication's efficiency showed 1 ml once-daily ante cibum oral administration to be the safest and most efficient regimen.
- The above method for obtaining individual fractions of Laminaria Japonica and/or Laminaria Angustata consisting of 5 to 10 kD molecules seems optimal and permits more than 10,000-fold enrichment of the fraction from its initial content.
- The proposed method's algorithm is as follows:
- To obtain the said fractions from dried and powdered homogenate of algae (Laminaria Japonica and Laminaria Angustata) by leaching with butanol and water mixture, we leach powdered Laminaria in water and isobutanol solution (1 g:2.5 ml:2.5 ml). To this end, we pour 500 ml of isobutanol into a 121 bottle, then add 200 g of powdered Laminaria and stir until the solution is homogeneous and dark green in colour. Ten minutes later, we add 500 ml of distilled water into the bottle and stir again until homogeneous. The resultant mixture is heated in water bath or a thermostat at 60° C. and then incubated for 10-15 hours at room temperature; then supernatant is withdrawn and centrifuged in an Eppendorf Centrifuge 5804 R at 3000 rpm and +4° C. for 30 minutes. The supernatant thus obtained should be withdrawn, placed into a vacuum evaporator and blown through with argon. The resultant supernatant is then vacuum-dried as follows: at room temperature for one hour, to avoid ebullition as suction is applied, and then at 60° C. until complete evaporation. Then we prepare 5% aqueous solution and ultracentrifuge it at 100,000 rpm for one hour. The resulting supernatant should be withdrawn and centrifuged again, now in 10 kD filter tubes, for one hour at 5000 rpm on the Eppendorf Centrifuge 5804 R apparatus. The lower percolated fraction is then withdrawn and centrifuged in 5 kD filter tubes, for one hour at 5000 rpm on the Eppendorf Centrifuge 5804 R7 apparatus. Then the upper fraction detained by the filter is withdrawn and 500 ml of NaCl saline is added. The medication obtained is used as follows: 1 ml of the medication is diluted in 20 ml of water and ingested once a day before a meal for two weeks. The medication consists of 5 to 10 kD molecules.
- A female patient, aged 40, was referred to an immunologist by her GP as she had had frequent ARD episodes (4-5 per year) in the past three years and complained of fatigue, drowsiness and persistent weakness. Her history was unburdened, with two pregnancies, both delivered (at the age of 23 and 27); physical examination found no abnormalities. The blood cell count and chemistry tests and clinical urine test found none, either. The immunologist ordered an examination of her immune status, which found a reduced pool of natural killers and B lymphocites while their absolute values remained normal. The patient was started on our medication, with 1 ml administered orally in 20 ml of water before meals, once daily for two weeks. One month after the start of treatment, the patient noted a subjective improvement in her status, with lesser weakness and fatigue; she found it easier to get up in the morning (while her daily regimen remained the same). Objectively, re-examination of her immune status showed an increase in her natural killer and B lymphocyte pools that doubled and trebled, respectively.
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Value found, % Norm Before One % Indicator the course month after (for cells) Absolute T lymphocytes 68 66 66-76 1.4-2*10{circumflex over ( )}9 (CD3+) B lymphocytes 5 10 12-22 0.3-0.5*10{circumflex over ( )}9 (CD19+) T helpers (CD3+ 39 35 33-41 0.7-1.1*10{circumflex over ( )}9 CD4+) Cytotoxic T cells 29 33 27-35 0.6-0.9*10{circumflex over ( )}9 (CD3+, CD8+) Natural killers 2 6 4-27 0.1-0.5*10{circumflex over ( )}9 (CD3+, CD16+, CD56+) CD4/CD8 1.3 1.3 1-1.5 Ig A, mg/ml 2.0 2.5 1.03-4.61 - A male patient aged 60, with no abnormal history, consulted the immunologist on his own and reported occasional joint pains, acrimony and sleeping problems (difficulty falling asleep and awakening). Examination at the internal medicine department of a private clinic found no abnormalities. Immune status examination showed decreased count of natural killers (relatively) and humoral immunity factors (IgA and IgM). The patient was started on our medication, with 1 ml administered orally in 20 ml of water before meals, once daily for two weeks. One month after start of treatment, the patient reported better sleep (quick going to sleep with no waking in the night) and improved mood. Noted objectively one month after the start of treatment were an increase in humoral immunity factors and normalized lymphocyte population ratios.
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Value found, % Norm Before One % Indicator the course month after (for cells) Absolute T lymphocytes 75 76 66-76 1.4-2*10{circumflex over ( )}9 (CD3+) B lymphocytes 15 17 12-22 0.3-0.5*10{circumflex over ( )}9 (CD19+) T helpers (CD3+ 32 32 33-41 0.7-1.1*10{circumflex over ( )}9 CD4+) Cytotoxic T cells 31 29 27-35 0.6-0.9*10{circumflex over ( )}9 (CD3+, CD8+) Natural killers 3.5 10 4-27 0.1-0.5*10{circumflex over ( )}9 (CD3+, CD16+, CD56+) CD4/CD8 1.3 1.3 1-1.5 Ig A, mg/ml 0.5 2.7 1.03-4.61 Ig G, mg/ml 7.5 8 6.2-14.7 Ig M, mg/ml 0.3 0.51 0.61-1.64 - The above examples show that the medication whose active substance is 5-10 kD molecules from Laminaria Japonica and Laminaria Angustata leads to normalization of the patients' immune status and a subjective feeling of improved health.
Claims (5)
1. A method for stimulating cells in the human immune system of a patient, comprising administering to the patient a substance prepared according to a method comprising:
leaching homogenate of Laminaria Japonica and Laminaria Angustata algae with a mixture of an aqueous solution and an isobutanolic solution, with a 1 g:2.5 ml:2.5 ml mix ratio, to obtain a homogeneous mixture;
heating the homogenous mixture at 60° C. for one hour;
incubating the homogenous mixture for 10-15 hours at room temperature to obtain a supernatant;
withdrawing the supernatant from the homogenous mixture and centrifuging the supernatant at 3000 rpm and +4° C. for 30 minutes to obtain a centrifuged supernatant;
purging the centrifuged supernatant with argon to obtain a purged,
centrifuged supernatant and vacuum drying the purged, centrifuged supernatant for one hour at room temperature, followed by vacuum drying at 60° C. until a liquid fraction of the supernatant evaporates, to obtain a sediment;
diluting the sediment with water to produce a 5% solution and then ultracentrifuging the 5% solution at 100,000 rpm for one hour to obtain a second supernatant;
withdrawing the second supernatant and centrifuging the second supernatant in 10 kD filter tubes for one hour at 5000 rpm to obtain a lower percolated fraction;
withdrawing the lower percolated fraction and centrifuging the lower percolated fraction in 5 kD filter tubes for one hour at 5000 rpm to obtain an upper fraction detained by a filter in the filter tubes; and
withdrawing the upper fraction from the filter and diluting the upper fraction in 500 ml of NaCl saline to obtain a substance with a final concentration of 10 mg/ml.
2. The method according to claim 1 , wherein the method for preparing the substance administered to the patient further comprises, after centrifuging the supernatant and prior to purging the centrifuged supernatant, ultracentrifuging the centrifuged supernatant at 100,000 rpm for one hour.
3. The method of claim 1 , wherein the substance is administered orally to the patient for a fortnight.
4. The method of claim 3 , wherein the substance is administered in 1 ml dosage once daily for a fortnight.
5. The method of claim 3 , wherein the substance is administered before meals.
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US17/002,169 US11534469B2 (en) | 2020-08-25 | 2020-08-25 | Method for the stimulation of human immune cells |
US17/993,633 US20230091957A1 (en) | 2020-08-25 | 2022-11-23 | Method for the Stimulation of Human Immune Cells |
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Citations (4)
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KR20100136739A (en) * | 2009-06-19 | 2010-12-29 | 인제대학교 산학협력단 | Composition for antioxidation and preventing or treating immune disease comprising laminaria japonica extract as an active ingredient |
KR101193558B1 (en) * | 2010-04-21 | 2012-10-22 | 부경대학교 산학협력단 | An anti-inflammatory pharmaceutical composition comprising of extracts as an effective component from Laminaria japonica |
WO2015020567A1 (en) * | 2013-08-06 | 2015-02-12 | Limited Liability Company "Nearmedic Plus" | Laminaria angustata-based product for enhancing regeneration and proliferation of cells, method of producing and use the same |
US11534469B2 (en) * | 2020-08-25 | 2022-12-27 | Vita Motus AG | Method for the stimulation of human immune cells |
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WO2011105431A1 (en) * | 2010-02-24 | 2011-09-01 | 森永乳業株式会社 | Antibacterial auxiliary agent comprising kombu extract as active ingredient, antibacterial composition, and food or beverage |
US10590207B2 (en) * | 2010-11-23 | 2020-03-17 | DuPont Nutrition USA, Inc. | Process for isolating fucoidan and laminarin from live, harvested seaweed |
RU2498821C1 (en) | 2012-04-10 | 2013-11-20 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Новосибирский национальный исследовательский государственный университет" (Новосибирский государственный университет, НГУ) | Method for stimulating endogenous production of cytokines and hemopoietins |
CL2015000876A1 (en) * | 2015-04-08 | 2015-08-28 | Maqui New Life S A | Veterinary composition of seaweed extract with at least 5% of fucoidians and andrographis sp plant extract with at least 5% of andrographolides, useful in the control and prevention of infections caused by intracellular microorganisms in fish. |
RU2717672C1 (en) | 2019-04-05 | 2020-03-25 | Гордейчук Владимир Евгеньевич | Method of human immune system cells stimulation |
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KR20100136739A (en) * | 2009-06-19 | 2010-12-29 | 인제대학교 산학협력단 | Composition for antioxidation and preventing or treating immune disease comprising laminaria japonica extract as an active ingredient |
KR101193558B1 (en) * | 2010-04-21 | 2012-10-22 | 부경대학교 산학협력단 | An anti-inflammatory pharmaceutical composition comprising of extracts as an effective component from Laminaria japonica |
WO2015020567A1 (en) * | 2013-08-06 | 2015-02-12 | Limited Liability Company "Nearmedic Plus" | Laminaria angustata-based product for enhancing regeneration and proliferation of cells, method of producing and use the same |
US11534469B2 (en) * | 2020-08-25 | 2022-12-27 | Vita Motus AG | Method for the stimulation of human immune cells |
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