WO1998058655A1 - Use of a humic acid-containing substance in medicine - Google Patents
Use of a humic acid-containing substance in medicine Download PDFInfo
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- WO1998058655A1 WO1998058655A1 PCT/HU1998/000059 HU9800059W WO9858655A1 WO 1998058655 A1 WO1998058655 A1 WO 1998058655A1 HU 9800059 W HU9800059 W HU 9800059W WO 9858655 A1 WO9858655 A1 WO 9858655A1
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- WIPO (PCT)
- Prior art keywords
- peat
- humic acid
- containing substance
- origin
- active ingredient
- Prior art date
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- 239000004021 humic acid Substances 0.000 title claims abstract description 63
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 239000000126 substance Substances 0.000 title claims abstract description 33
- 239000003814 drug Substances 0.000 title abstract description 3
- 239000003415 peat Substances 0.000 claims abstract description 31
- 239000000725 suspension Substances 0.000 claims abstract description 17
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 13
- 230000008569 process Effects 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 10
- 235000017913 Scirpus lacustris Nutrition 0.000 claims abstract description 8
- 239000004480 active ingredient Substances 0.000 claims abstract description 8
- 239000012670 alkaline solution Substances 0.000 claims abstract description 8
- 239000007858 starting material Substances 0.000 claims abstract description 7
- 241000196324 Embryophyta Species 0.000 claims abstract description 6
- 235000008573 Scirpus validus Nutrition 0.000 claims abstract description 5
- 239000007787 solid Substances 0.000 claims abstract description 5
- 238000003756 stirring Methods 0.000 claims abstract description 5
- 239000000654 additive Substances 0.000 claims abstract description 4
- 230000018044 dehydration Effects 0.000 claims abstract description 4
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 4
- 230000000366 juvenile effect Effects 0.000 claims abstract description 3
- 239000000969 carrier Substances 0.000 claims abstract 2
- 230000003394 haemopoietic effect Effects 0.000 claims description 18
- 230000008929 regeneration Effects 0.000 claims description 13
- 238000011069 regeneration method Methods 0.000 claims description 13
- 230000006378 damage Effects 0.000 claims description 9
- 244000078283 Scirpus lacustris Species 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 241000985128 Cladium mariscus Species 0.000 claims description 3
- 230000000069 prophylactic effect Effects 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 159000000011 group IA salts Chemical class 0.000 claims description 2
- 229910000000 metal hydroxide Inorganic materials 0.000 claims description 2
- 150000004692 metal hydroxides Chemical class 0.000 claims description 2
- 238000011321 prophylaxis Methods 0.000 claims 1
- 230000001502 supplementing effect Effects 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 244000143231 Scirpus validus Species 0.000 abstract 1
- 241001465754 Metazoa Species 0.000 description 32
- 239000000203 mixture Substances 0.000 description 24
- 238000011282 treatment Methods 0.000 description 15
- 239000000463 material Substances 0.000 description 10
- 210000000265 leukocyte Anatomy 0.000 description 8
- 231100000054 whole-body exposure Toxicity 0.000 description 8
- 241000700159 Rattus Species 0.000 description 6
- 208000027418 Wounds and injury Diseases 0.000 description 6
- 230000004071 biological effect Effects 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 208000014674 injury Diseases 0.000 description 6
- 238000010171 animal model Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000001035 drying Methods 0.000 description 4
- 239000008187 granular material Substances 0.000 description 4
- RYCLIXPGLDDLTM-UHFFFAOYSA-J tetrapotassium;phosphonato phosphate Chemical compound [K+].[K+].[K+].[K+].[O-]P([O-])(=O)OP([O-])([O-])=O RYCLIXPGLDDLTM-UHFFFAOYSA-J 0.000 description 4
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000000227 grinding Methods 0.000 description 3
- 230000002489 hematologic effect Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 229920002261 Corn starch Polymers 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 235000019759 Maize starch Nutrition 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 238000000889 atomisation Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000005469 granulation Methods 0.000 description 2
- 230000003179 granulation Effects 0.000 description 2
- 238000005534 hematocrit Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000005865 ionizing radiation Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000001995 reticulocyte Anatomy 0.000 description 2
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000008133 Iron-Binding Proteins Human genes 0.000 description 1
- 108010035210 Iron-Binding Proteins Proteins 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- GDCRSXZBSIRSFR-UHFFFAOYSA-N ethyl prop-2-enoate;2-methylprop-2-enoic acid Chemical compound CC(=C)C(O)=O.CCOC(=O)C=C GDCRSXZBSIRSFR-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000009969 flowable effect Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000013383 initial experiment Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000011418 maintenance treatment Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000000191 radiation effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 235000000891 standard diet Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 229910000406 trisodium phosphate Inorganic materials 0.000 description 1
- 235000019801 trisodium phosphate Nutrition 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/02—Medicinal preparations containing materials or reaction products thereof with undetermined constitution from inanimate materials
- A61K35/10—Peat; Amber; Turf; Humus
Definitions
- the invention relates to the use of a humic acid-containing substance in medicine, to pharmaceutical compositions containing as an active ingredient such a substance, and to a process for preparing this substance.
- the pharmaceutical compositions are useful as a prophylactic to injuries of the haemopoietic system or for regeneration of the injured haemopoietic system.
- humic acids are heteropolycondensates of widely various compositions, respectively, allomelanins, which can be found, e.g. in soils, carbon sorts and peat. Humic acids are formed through a slow decomposition process, respectively chemical and biological transformation of plant materials.
- W. Flaig "The chemistry of humic substances", FAO/IAEA Tech. Meet. Brunswick- V ⁇ lkenrode, p. 103 to 127 (1963); . V. Cheshire et al.: “Humic Acids II. Structure of humic acids", Tetrahedron, 23, 1669-1682 (1967); M. Schnitzer and S. U.
- humic acids contain complex, polymerized macromolecules of phenolic structure; their composition depends strongly on the site and age of their formation.
- the metal ion- binding, particularly iron-binding, and chelate-forming properties of humic acids are well known.
- the main peat-forming plants of the starting material referred to above are 20 to 40% of great bulrush (Schoenoplectus lacustris) and 60 to 80% of winter-sedge (Cladium mariscus).
- the biological effects of these humic acid-containing substances were studied in several series of experiments. It surprisingly has been found that the humic acid-containing substances prepared by the process to be described later favourably influence the regeneration of the haemopoietic system injured by external whole body 60 Co gamma irradiation of the experimental animals.
- mice Female Wistar rats of 190 to 220 g body-weight (Laboratory Animals Institute, G ⁇ d ⁇ ll ⁇ , Hungary), randomized according to weight, were used in test groups. The animals were kept in plastic cages (5 animals in each cage) at a controlled temperature of 23 ⁇ 3 °C and relative humidity of 60 ⁇ 10% under alternating illumination (12-hour cycles of light/darkness). The rats received standard diet (Code: 624, Altromin GmbH, D-32791 Heil/Lippe, Germany) and water ad libitum. The average daily food consumption was 20 g per animal.
- the rats were acclimated to the experimental conditions for two weeks. During the experiment, the general physical condition of the animals was controlled daily. II. Test material
- the preparations were administered to the experimental animals in various doses through a gastric tube.
- Natural humic acid product for feeding (in the following text: HA) was prepared by grinding the normal standard rat pellet (see earlier) mixed with the required amount of the material as prepared according to the following Example 1. The mixture was homogenized, regranulated and dried at room temperature.
- Gray Gray
- LD50/30 value characteristic of the rat strain was found to be 7.5 Gy.
- the haematological parameters including white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin (HGB), haematocrit (HTC), platelet (PLT) count, and reticulocyte (RET) count, were determined by using MEDICOR PHA-1 and PHA-2 type haematologic automatic devices (MEDICOR Ltd., Budapest, Hungary).
- Group 1 Whole body irradiated with 7 Gy of 60 Co gamma (standard feed + tap water).
- Group 2 Pretreatment with HA for 7 days (240 mg/animal/day), then whole body exposure to 7 Gy and additional 4-week treatment with a dose of 240 mg/animal/day of HA.
- Group 3 Whole-body exposure to 7 Gy, then a single treatment with a dose of 240 mg/animal/day of HA.
- Group 4 Pretreatment with HA for 7 days (90 mg/animal/day) then whole- body exposure to 7 Gy and additional 4-week treatment with a dose of 90 mg/animal/day of HA.
- Group 5 Whole-body exposure to 7 Gy, then a single treatment with a dose of 90 mg/animal/day of HA.
- the experimental data were analyzed by using the Student's T test.
- test material applied exerts its effect in several therapeutical doses in the case of a high-dose 60 Co gamma whole-body exposure in the normalization of the irradiation- induced injuries of the haemopoietic system.
- the action becomes most favourable by also using the test material before irradiation as a pretreatment.
- humic acid- containing compositions according to the invention can be used advantageously to prevent injuries to the haemopoietic system of various origins or for an effective, increased regeneration of the injured haemopoietic system.
- compositions according to the present invention can be effectively used in human therapy in cases in which the human body is affected by an ionizing radiation during therapy or because of other events (reactor accident, accidental radiation effect affecting the patient or the handling crew, etc.). Additional experiments indicate that the compositions according to the present invention can also be utilized to increase the regeneration of the haemopoietic system when it is injured during chemotherapy treatments.
- the invention relates to a pharmaceutical composition useful for the prevention of injuries of the haemopoietic system and for regeneration of an injured haemopoietic system, in which the composition comprises an effective amount, suitably 0.01 to 99.9% by weight, of humic acid-containing substance of peat origin together with the usual additives.
- the compositions contain 1.0 to 25.0% by weight of humic acid-containing substance of fen peat origin and 75 to 99% by weight of known, therapeutical ly acceptable additive(s) in solid, liquid, or gel form.
- the invention further relates to a process for the preparation of a humic acid-containing substance.
- This process is characterized in that, as a starting material, juvenile peat of fen origin of not more than 15,000 years of age is used, which had been formed from at least 20% of great bulrush (Schoenoplectus lacustris) and at least 40% of winter-sedge (Cladium mariscus) as peat-forming plants.
- the peat is stirred with an aqueous alkaline solution of pH 7.5 to 10.5 to obtain a suspension.
- at least a 2.5-fold amount of an alkaline solution is used to form the suspension.
- An aqueous solution of alkaline metal hydroxides preferably sodium or potassium hydroxide, and/or a basic aqueous solution of alkaline salts, like sodium carbonate, trisodium phosphate, or potassium pyrophosphate, may be used as the aqueous alkaline solution.
- the upper phase obtained after settling of the mixture is suitably adjusted to 10 to 100 g/L of humic acid content, or concentrated by dehydration, or brought into solid form. Evaporation under reduced pressure, drying with atomization, lyophilization, and other or similar technical procedures may be applied.
- the appropriately selected peat is homogenized by drying, crushing, and grinding; the peat grist containing humic acid is suspended in a mildly alkaline, i.e. pH 7.5 to 10.5, aqueous medium, and settled; after separation from the lower phase, the pH value of the supernatant is, if desired, adjusted to pH 5 to 7, and then, if desired, the mixture is concentrated or dried by dehydration.
- a mildly alkaline i.e. pH 7.5 to 10.5, aqueous medium, and settled
- the pH value of the supernatant is, if desired, adjusted to pH 5 to 7, and then, if desired, the mixture is concentrated or dried by dehydration.
- the appropriately selected dried and homogenized peat is treated with a dilute, e.g. at most 5% by weight, alkaline solution of pH 7.5 to 10.5; then, the dry substance content of the recovered suspension is determined, and the humic acid-containing substance is transformed into the desired form, e.g. suspended or powdered.
- a dilute e.g. at most 5% by weight, alkaline solution of pH 7.5 to 10.5; then, the dry substance content of the recovered suspension is determined, and the humic acid-containing substance is transformed into the desired form, e.g. suspended or powdered.
- the advantages of the invention can be summarized as follows: 1 )
- the pharmaceutical compositions according to the present invention act within a novel and highly significant field by curing diseases related to injuries of the haemopoietic system; and 2)
- the invention provides a well-reproducible process for the preparation of the pharmaceutical compositions by ensuring a humic acid-containing substance of stable composition.
- Peat of 3,000 to 7,000 years of age originating from a depth of 0.5 to 2.5 m was used as the starting material, containing 20 to 40% of great bulrush and 60 to 80% of winter-sedge as peat-forming plants.
- the peat was treated with a 1 % potassium pyrophosphate (K4P2O7) solution at 45°C in an acid-resistant or enamel-lined vessel, fitted with a heater and a stirrer, under constant stirring. After dissolving 1 part by weight of potassium pyrophosphate in 100 parts by weight of tap water heated to a maximum of 45°C, 50 parts by weight of peat having a maximum moisture content of 30% were added. The suspension obtained contained about 10% of peat material. The recovery lasted not more than 48 hours. Within this period, after stirring for 1 hour, the mixture was subjected to vigorous, particle-grinding stirring for an additional 2 hours in a Dispax reactor (IKA Werke, Germany) for achieving a homogeneous particle distribution.
- K4P2O7 potassium pyrophosphate
- the humic acid-containing liquid removed from the settling tank by suction was stirred in the dilution tank for at least 30 minutes.
- 30 ml of homogeneous humic acid- containing suspension were dried to constant weight at 130°C in a drying oven.
- Example 3 After adding 1400 g of maize starch and 70 g of poly vinyl pyrrolidone to 1 L of a suspension according to Example 1 , containing 30 g/L of humic acid, a granulation mixture was prepared, granulated through a screen of 1.2 mm, dried at room temperature, i.e. maximum at 25°C, and re- granulated to give grayish loose granules.
- humic acid granules from a concentrate
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to the use of a humic acid-containing substance in medicine, to pharmaceutical compositions containing as active ingredient such a substance, and to a process for preparing this substance. The humic acid-containing substance according to the invention is preferably used in the form of a pharmaceutical composition containing 0.01 to 99.90 % by weight of humic acid-containing substance of peat origin as active ingredient together with carriers and/or other known additives. The invention also relates to a process for the preparation of a humic acid-containing substance of peat origin, which comprises using as the starting material a juvenile, preferably at most 10,000 years old, peat formed from a great bulrush and winter-sedge as peat-forming plants; preparing a suspension by stirring the peat with an aqueous alkaline solution of pH 7.5 to 10.5; utilizing the upper phase of the obtained suspension by adjusting its humic acid content to 10-100 g/L; concentrating it; or bringing it into solid form by dehydration.
Description
USE OF A HUMIC ACID-CONTAINING SUBSTANCE IN MEDICINE
The invention relates to the use of a humic acid-containing substance in medicine, to pharmaceutical compositions containing as an active ingredient such a substance, and to a process for preparing this substance. The pharmaceutical compositions are useful as a prophylactic to injuries of the haemopoietic system or for regeneration of the injured haemopoietic system.
It is known that humic acids are heteropolycondensates of widely various compositions, respectively, allomelanins, which can be found, e.g. in soils, carbon sorts and peat. Humic acids are formed through a slow decomposition process, respectively chemical and biological transformation of plant materials. [For details, see, e.g., W. Flaig: "The chemistry of humic substances", FAO/IAEA Tech. Meet. Brunswick- Vδlkenrode, p. 103 to 127 (1963); . V. Cheshire et al.: "Humic Acids II. Structure of humic acids", Tetrahedron, 23, 1669-1682 (1967); M. Schnitzer and S. U. Khan: "Humic substances in the environment", Dekker, New York (1972); C. Steelinek: "What is Humic Acid?", J. Chem. Educ, 40, 379-384 (1963)]. The humic acids contain complex, polymerized macromolecules of phenolic structure; their composition depends strongly on the site and age of their formation. The metal ion- binding, particularly iron-binding, and chelate-forming properties of humic acids are well known.
Recognition of such pioneering character is disclosed in the Hungarian patent specification No. 158,252, which describes a
composition for satisfying the trace element demand of vertebrates. This composition contains at least 8% of humic acid and metal ions, being in a biologically available form for vertebrates.
Since the end of the 80's, more and more publications devoted to the biological effects of humic acids or humic acid-containing materials have appeared. Thus, the antiallergic effect (German patent specification No. 4,335,523) and antiviral action (German patent specification No. 4,134,378) of humic acids and their salts became known; in addition, within the category of their antiviral action, the humic acids also were found to be useful for the treatment of HIV infections (see the international patent application No. WO 95/08335). Furthermore, data regarding the dermatological, bactericidal, gastrointestinal, and anti-inflammatory effects of humic acids also can be found [for details, see, e.g., Cr. Heinrich: "Huminsaure und Permeabilitat", Protoplasma 58, 402-425 (1964); R. Klδcking et al.: "Zur Biochemie der Huminsauren V. Die Bindung der Huminsauren an Serumproteine in vitro", Acta Biol. Med. German, 18, 9-13 (1967); R. Obenaus and R. Mϋcke: "Zur Biochemie der Huminsauren aus ihren Eisenchelatverbindungen", Acta Biol. Med. German, 10, 233-238 (1963)]. Our initial experiments have been directed to produce a reproducible, standardized humic acid mixture which can be used in the preparation of therapeutical compositions. Thus, our aim was to provide a humic acid mixture obtained by recovery and extraction of a peat originating from a well-characterized source, said peat produced and pre-treated under controlled conditions according to a uniform technology.
It has been found, on the basis of data and recoveries taken from various peat areas, that a standardized base material suitable for the preparation of pharmaceutical compositions can only be achieved by selecting a peat of not more than 15,000 years old, according to
radiocarbon determinations, as the starting material. A homogeneous peat layer of between 3,000 and 7,000 years of age (based on radiocarbon data) and found at depth of between 0.5 to 2.5 m under the ground surface proved to be particularly advantageous. The fibrous structure of this material can be well perceived by the naked eye.
According to the palaeobotanic data, the main peat-forming plants of the starting material referred to above are 20 to 40% of great bulrush (Schoenoplectus lacustris) and 60 to 80% of winter-sedge (Cladium mariscus). The biological effects of these humic acid-containing substances were studied in several series of experiments. It surprisingly has been found that the humic acid-containing substances prepared by the process to be described later favourably influence the regeneration of the haemopoietic system injured by external whole body 60Co gamma irradiation of the experimental animals.
This recognition is novel and original since no literature data or references were found which would prove such biological activity of humic acid.
Detailed investigations were carried out on experimental animals by using various doses of whole-body gamma irradiation in order to develop a composition and a method of treatment that can be effectively used for human therapy as well.
The biological activity of humic acid-containing substances prepared by the process of the present invention was proven by the experimental results described below.
I. Experimental animals
At the beginning of the experiments, female Wistar rats of 190 to 220 g body-weight (Laboratory Animals Institute, Gόdόllδ, Hungary), randomized according to weight, were used in test groups. The animals
were kept in plastic cages (5 animals in each cage) at a controlled temperature of 23 ± 3 °C and relative humidity of 60 ± 10% under alternating illumination (12-hour cycles of light/darkness). The rats received standard diet (Code: 624, Altromin GmbH, D-32791 Lage/Lippe, Germany) and water ad libitum. The average daily food consumption was 20 g per animal.
The rats were acclimated to the experimental conditions for two weeks. During the experiment, the general physical condition of the animals was controlled daily. II. Test material
The preparations were administered to the experimental animals in various doses through a gastric tube.
Natural humic acid product for feeding (in the following text: HA) was prepared by grinding the normal standard rat pellet (see earlier) mixed with the required amount of the material as prepared according to the following Example 1. The mixture was homogenized, regranulated and dried at room temperature.
III. 60co gamma whole body irradiation
The whole body exposure of rats was performed in a special plastic cage (40 animals/cage) with an irradiation dose of 7.0 Gray (Gy) (dose intensity = 0.82 Gy/min). The LD50/30 value characteristic of the rat strain was found to be 7.5 Gy.
IV. Haematoloqical investigations
On days 0, 7, 14, 21 and 28 of the experiment, the animals were anaesthetized by ether; the abdominal section of the aorta was prepared and blood samples were taken.
The haematological parameters, including white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin (HGB), haematocrit (HTC), platelet (PLT) count, and reticulocyte (RET) count, were
determined by using MEDICOR PHA-1 and PHA-2 type haematologic automatic devices (MEDICOR Ltd., Budapest, Hungary).
V. Experimental groups
In our experiments, treatments were performed on 30 animals in each group with various doses of the natural humic acid (HA) product as defined in point II above. The mean haematological values characteristic of the WISTAR strain are shown as reference values in the Figures in every case.
Group 1 : Whole body irradiated with 7 Gy of 60Co gamma (standard feed + tap water).
Group 2: Pretreatment with HA for 7 days (240 mg/animal/day), then whole body exposure to 7 Gy and additional 4-week treatment with a dose of 240 mg/animal/day of HA. Group 3: Whole-body exposure to 7 Gy, then a single treatment with a dose of 240 mg/animal/day of HA.
Group 4: Pretreatment with HA for 7 days (90 mg/animal/day) then whole- body exposure to 7 Gy and additional 4-week treatment with a dose of 90 mg/animal/day of HA. Group 5: Whole-body exposure to 7 Gy, then a single treatment with a dose of 90 mg/animal/day of HA.
The experimental data were analyzed by using the Student's T test.
VI. Results
From the haematological parameters of the rats treated as above, the changes in WBC count and PLT count are shown in Figures 1 and 2 for the treatment with a dose of 240 mg/animal/day and in Figures 3 and 4 for the treatment with a dose of 90 mg/animal/day. In the Figures, the number of days post-exposure are indicated on the horizontal axes, whereas the cell counts as G/L (10 E 09/L) values are given on the vertical axes.
Results obtained bv treatment with a dose of 240 mg/animal/dav
It can be stated that the WBC and PLT counts were significantly (p <
0.05) decreased in the whole-body irradiated animals (group 1 , control) as well as in animals treated once with HA after irradiation (group 3). A medium-grade enhancement of regeneration caused by a single HA treatment occurred only with respect to PLT starting from the 3rd week.
The regeneration of both the WBCs and PLTs of animals in the untreated control group (group 1) began only after the 3rd week following the irradiation, No injury of the haemopoietic system occurred in the animals pretreated with 240 mg/animal of HA and then additionally treated with the same dose of HA after whole-body exposure to 7 Gy (group 2), and the number of WBCs and PLTs remained near the reference values characteristic for the WISTAR strain, These experimental results demonstrate that the harmful biological effect of a high dose of ionizing radiation on the haemopoietic system practically can be avoided by administering a suitable dose (240 mg/animal/day) of the test material and by using a proper method of treatment [treatment preceding the irradiation, followed by a maintenance treatment] (c.f. Figures 1 and 2).
Results obtained bv treatment with a dose of 90 mg/animal/dav It can be stated that both the WBC and PLT counts were significantly (p < 0.05) decreased in the case of either the once-treated or the continuously treated animals by one week following the whole-body exposure. A significantly decreased PLT count was measured in the animals both in the untreated control group (group 1) and also in the animals treated once with HA (group 5) also in the 2nd week post- irradiation.
The regeneration of WBCs as well as PLTs began in the animals in
WO 98/58655 ~ ' " PCT/HU98/00059 the irradiated but untreated control group (group 1) only after the 3rd week following irradiation.
When pretreatment with HA was carried out on animals treated with a dose of 90 mg/animal/day of HA (group 4), the regeneration of both cell types already had started intensively after the first week and reached the values of the control animals by the end of the 2nd week (Figures 3 and 4).
A regeneration to a similar degree as that of the pretreated group (alteration of the PLT, Figure 4) occurred in the group treated once with a dose of 90 mg/animal/day (group 5) only from the 2nd week, In summary, it can be stated that the test material applied exerts its effect in several therapeutical doses in the case of a high-dose 60Co gamma whole-body exposure in the normalization of the irradiation- induced injuries of the haemopoietic system. The action becomes most favourable by also using the test material before irradiation as a pretreatment.
It is unambiguously proven by the above results that the humic acid- containing compositions according to the invention can be used advantageously to prevent injuries to the haemopoietic system of various origins or for an effective, increased regeneration of the injured haemopoietic system.
Due to their proven biological activity, the compositions according to the present invention can be effectively used in human therapy in cases in which the human body is affected by an ionizing radiation during therapy or because of other events (reactor accident, accidental radiation effect affecting the patient or the handling crew, etc.). Additional experiments indicate that the compositions according to the present invention can also be utilized to increase the regeneration of the haemopoietic system when it is injured during chemotherapy treatments.
Thus, the invention relates to a pharmaceutical composition useful
for the prevention of injuries of the haemopoietic system and for regeneration of an injured haemopoietic system, in which the composition comprises an effective amount, suitably 0.01 to 99.9% by weight, of humic acid-containing substance of peat origin together with the usual additives. According to a preferred embodiment of the invention, the compositions contain 1.0 to 25.0% by weight of humic acid-containing substance of fen peat origin and 75 to 99% by weight of known, therapeutical ly acceptable additive(s) in solid, liquid, or gel form.
The invention further relates to a process for the preparation of a humic acid-containing substance. This process is characterized in that, as a starting material, juvenile peat of fen origin of not more than 15,000 years of age is used, which had been formed from at least 20% of great bulrush (Schoenoplectus lacustris) and at least 40% of winter-sedge (Cladium mariscus) as peat-forming plants. The peat is stirred with an aqueous alkaline solution of pH 7.5 to 10.5 to obtain a suspension. Suitably, at least a 2.5-fold amount of an alkaline solution, based on the weight of the peat, is used to form the suspension. An aqueous solution of alkaline metal hydroxides, preferably sodium or potassium hydroxide, and/or a basic aqueous solution of alkaline salts, like sodium carbonate, trisodium phosphate, or potassium pyrophosphate, may be used as the aqueous alkaline solution. The upper phase obtained after settling of the mixture is suitably adjusted to 10 to 100 g/L of humic acid content, or concentrated by dehydration, or brought into solid form. Evaporation under reduced pressure, drying with atomization, lyophilization, and other or similar technical procedures may be applied.
In more detail, the appropriately selected peat is homogenized by drying, crushing, and grinding; the peat grist containing humic acid is suspended in a mildly alkaline, i.e. pH 7.5 to 10.5, aqueous medium, and settled; after separation from the lower phase, the pH value of the
supernatant is, if desired, adjusted to pH 5 to 7, and then, if desired, the mixture is concentrated or dried by dehydration.
According to a preferred embodiment of the process of the present invention, the appropriately selected dried and homogenized peat is treated with a dilute, e.g. at most 5% by weight, alkaline solution of pH 7.5 to 10.5; then, the dry substance content of the recovered suspension is determined, and the humic acid-containing substance is transformed into the desired form, e.g. suspended or powdered.
The advantages of the invention can be summarized as follows: 1 ) The pharmaceutical compositions according to the present invention act within a novel and highly significant field by curing diseases related to injuries of the haemopoietic system; and 2) The invention provides a well-reproducible process for the preparation of the pharmaceutical compositions by ensuring a humic acid-containing substance of stable composition.
The invention is illustrated by the following non-limiting Examples.
Example 1
Production of a humic acid-containing substance
Peat of 3,000 to 7,000 years of age originating from a depth of 0.5 to 2.5 m was used as the starting material, containing 20 to 40% of great bulrush and 60 to 80% of winter-sedge as peat-forming plants.
The peat was treated with a 1 % potassium pyrophosphate (K4P2O7) solution at 45°C in an acid-resistant or enamel-lined vessel, fitted with a heater and a stirrer, under constant stirring. After dissolving 1 part by weight of potassium pyrophosphate in 100 parts by weight of tap water heated to a maximum of 45°C, 50 parts by weight of peat having a maximum moisture content of 30% were added. The suspension obtained contained about 10% of peat material. The recovery lasted not more than 48 hours. Within this period, after stirring for 1 hour, the mixture was
subjected to vigorous, particle-grinding stirring for an additional 2 hours in a Dispax reactor (IKA Werke, Germany) for achieving a homogeneous particle distribution.
After 48 hours, the suspension of recovered peat was pumped into a settling tank and settled for 24 hours. The upper phase was separated from the lower, solid sediment after 48 hours.
The humic acid-containing liquid removed from the settling tank by suction was stirred in the dilution tank for at least 30 minutes. In order to determine the dry substance content, 30 ml of homogeneous humic acid- containing suspension were dried to constant weight at 130°C in a drying oven. The suspension was adjusted to a humic acid content of 60 g/L concentration with a 1% potassium pyrophosphate solution (pH=9.4), based on the dry substance content determination.
Thus, a standardized humic acid suspension of 60 g/L concentration was obtained. Example 2
Preparation of humic acid granulates
After adding 1400 g of maize starch and 70 g of poly vinyl pyrrolidone to 1 L of a suspension according to Example 1 , containing 30 g/L of humic acid, a granulation mixture was prepared, granulated through a screen of 1.2 mm, dried at room temperature, i.e. maximum at 25°C, and re- granulated to give grayish loose granules. Example 3
Preparation of humic acid granules from a concentrate A mixture of 1 L of starting material employed in Example 2, i.e. the suspension containing preferably 30 g/L of humic acid, was evaporated down to a volume of 100 to 120 mL, and, after adding 11 g of polyvinylpyrrolidone and 110 g of maize starch to the suspension, a granulation mixture was prepared. This mixture was granulated through a
1.2 mm screen, dried at room temperature, i.e. maximum at 25°C, and re- granulated to give grayish granules.
Example 4
Preparation of a microcapsulated humic acid composition Four hundred grams of 30% Eudragit L 30 D dispersion (Rohm
Pharma, Darmstadt, Germany) were added to a suspension containing the substance of Example 1 in an amount of 100 g of dry substance, and mixed thoroughly. The mixture obtained was dried through atomization by using an inlet drying air flow of maximum 40°C and a feeding rate of 750 g/hour to give a fine, nearly black, flowable powder.
Claims
1. Use of a humic acid-containing substance of peat origin as a prophylactic to injuries of the haemopoietic system or for the regeneration of the injured haemopoietic system.
2. Use according to claim 1 in the form of a pharmaceutical composition comprising 0.01 to 99.90% by weight of humic acid- containing substance of peat origin as active ingredient together with, in an amount supplementing up to 100%, carriers and/or other known additives.
3. Use according to claim 1 or 2 in the form of a pharmaceutical composition containing 1.0 to 25.0% by weight of active ingredient.
4. A pharmaceutical composition useful for the prophylaxis of injuries of the haemopoietic system or for regeneration of the injured haemopoietic system, which comprises an effective amount of a humic acid-containing substance of peat origin as active ingredient.
5. A pharmaceutical composition according to claim 4, which comprises 0.01 to 99.90% by weight of active ingredient.
6. A pharmaceutical composition according to claim 4 or 5, which comprises 1.0 to 25.0% by weight of active ingredient.
7. Process for the preparation of a humic acid-containing substance of peat origin according to any of the preceding claims, which comprises using as the starting material a juvenile, preferably at most 10,000 years old, peat formed from at least 20% of great bulrush (Schoenoplectus lacustris) and at least 40% of winter-sedge (Cladium mariscus) as peat- forming plants; preparing a suspension by stirring the peat with an aqueous alkaline solution of 7.5 to 10.5 pH value; utilizing the upper phase of the suspension obtained after settling by adjusting its humic acid content to a value suitably between 10 and 100 g/L; concentrating it; or bringing it into solid form by dehydration.
8. A process according to claim 7, which comprises using an aqueous alkaline solution in an at least 2-5-fold amount, based on the peat weight, during formation of the suspension.
9. A process according to claim 7 or 8, which comprises using an aqueous solution of an alkaline metal hydroxide or a basic aqueous solution of an alkaline salt as aqueous alkaline solution.
10. Method for use as a prophylactic to injuries of the haemopoietic system or for the regeneration of the injured haemopoietic system, characterized by administering an effective amount of humic acid- containing substance of peat origin to the patient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU82362/98A AU8236298A (en) | 1997-06-24 | 1998-06-23 | Use of a humic acid-containing substance in medicine |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| HUP9701093 | 1997-06-24 | ||
| HU9701093A HUP9701093A1 (en) | 1997-06-24 | 1997-06-24 | Pharmaceutical composition containing huminic acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1998058655A1 true WO1998058655A1 (en) | 1998-12-30 |
Family
ID=89995281
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/HU1998/000059 WO1998058655A1 (en) | 1997-06-24 | 1998-06-23 | Use of a humic acid-containing substance in medicine |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU8236298A (en) |
| HU (1) | HUP9701093A1 (en) |
| WO (1) | WO1998058655A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002024609A2 (en) * | 2000-09-19 | 2002-03-28 | Nobel Limited Liability Company | Method for producing a radioprotector |
| EP1369122A1 (en) * | 2001-02-21 | 2003-12-10 | Nobel Limited Liability Company | Anticancer agent |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992016216A1 (en) * | 1991-03-16 | 1992-10-01 | Torf Establishment | Peat-derived bioactive products and pharmaceutical and cosmetic compositions containing them |
| WO1995008335A1 (en) * | 1993-09-24 | 1995-03-30 | Maurizio Zanetti | Treatment of hiv infection with humic acid |
-
1997
- 1997-06-24 HU HU9701093A patent/HUP9701093A1/en unknown
-
1998
- 1998-06-23 AU AU82362/98A patent/AU8236298A/en not_active Abandoned
- 1998-06-23 WO PCT/HU1998/000059 patent/WO1998058655A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992016216A1 (en) * | 1991-03-16 | 1992-10-01 | Torf Establishment | Peat-derived bioactive products and pharmaceutical and cosmetic compositions containing them |
| WO1995008335A1 (en) * | 1993-09-24 | 1995-03-30 | Maurizio Zanetti | Treatment of hiv infection with humic acid |
Non-Patent Citations (3)
| Title |
|---|
| DATABASE MEDLINE US NATIONAL LIBRARY OF MEDICINE (NLM), BETHESDA, MD, US; PUKHOVA G G ET AL: "[Effect of sodium humate on animals irradiated with lethal doses]. Vliianie gumata natriia na zhivotnykh, obluchennykh v letal'nykh dozakh.", XP002080637 * |
| INGLOT A D ET AL: "TOLPA TORF PREPARATION (TTP) INDUCES INTERFERON AND TUMOR NECROSIS FACTOR PRODUCTION IN HUMAN PERIPHERAL BLOOD LEUKOCYTES", ARCHIVUM IMMUNOLOGIAE ET THERAPIAE EXPERIMENTALIS, vol. 41, no. 1, 1993, pages 73 - 80, XP000619722 * |
| RADIOBIOLOGIIA, (1987 SEP-OCT) 27 (5) 650-3 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002024609A2 (en) * | 2000-09-19 | 2002-03-28 | Nobel Limited Liability Company | Method for producing a radioprotector |
| WO2002024609A3 (en) * | 2000-09-19 | 2002-08-15 | Nobel Ltd Liability Company | Method for producing a radioprotector |
| EP1369122A1 (en) * | 2001-02-21 | 2003-12-10 | Nobel Limited Liability Company | Anticancer agent |
| EP1369122A4 (en) * | 2001-02-21 | 2004-05-06 | Nobel Ltd Liability Company | Anticancer agent |
| EP1810684A2 (en) * | 2001-02-21 | 2007-07-25 | Nobel Limited Liability Company | Anticancer agent |
| EP1810684A3 (en) * | 2001-02-21 | 2008-04-16 | Nobel Limited Liability Company | Anticancer agent |
Also Published As
| Publication number | Publication date |
|---|---|
| HU9701093D0 (en) | 1997-08-28 |
| AU8236298A (en) | 1999-01-04 |
| HUP9701093A1 (en) | 1999-08-30 |
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