LV13366B - Standardised steroid saponin mixture, a method of its obtaining and application - Google Patents
Standardised steroid saponin mixture, a method of its obtaining and application Download PDFInfo
- Publication number
- LV13366B LV13366B LVP-05-96A LV050096A LV13366B LV 13366 B LV13366 B LV 13366B LV 050096 A LV050096 A LV 050096A LV 13366 B LV13366 B LV 13366B
- Authority
- LV
- Latvia
- Prior art keywords
- extraction
- butanol
- methanol
- mixture
- water
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 14
- 238000000034 method Methods 0.000 title claims abstract description 13
- 150000005856 steroid saponins Chemical class 0.000 title claims abstract description 9
- 229930182490 saponin Natural products 0.000 claims abstract description 12
- 150000007949 saponins Chemical class 0.000 claims abstract description 12
- 235000017709 saponins Nutrition 0.000 claims abstract description 12
- 241001533104 Tribulus terrestris Species 0.000 claims abstract description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- 238000000605 extraction Methods 0.000 claims description 16
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- 239000002021 butanolic extract Substances 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 5
- 239000012141 concentrate Substances 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 4
- 229940079593 drug Drugs 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- 230000002519 immonomodulatory effect Effects 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 230000003308 immunostimulating effect Effects 0.000 claims description 2
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 2
- 238000003786 synthesis reaction Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 239000008213 purified water Substances 0.000 claims 1
- 239000002955 immunomodulating agent Substances 0.000 abstract 1
- 229940121354 immunomodulator Drugs 0.000 abstract 1
- 230000002584 immunomodulator Effects 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 description 13
- 239000000463 material Substances 0.000 description 10
- 241000196324 Embryophyta Species 0.000 description 9
- 230000001681 protective effect Effects 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 230000003641 microbiacidal effect Effects 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000003306 harvesting Methods 0.000 description 4
- 230000000242 pagocytic effect Effects 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- LVTJOONKWUXEFR-UEZXSUPNSA-N protodioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@@H]5O[C@]([C@H]([C@@H]5[C@@]4(C)CC[C@@H]3[C@@]2(C)CC1)C)(O)CC[C@@H](C)CO[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O LVTJOONKWUXEFR-UEZXSUPNSA-N 0.000 description 3
- MHKGPHKABOLURA-JNVLQWCMSA-N protodioscin Natural products C[C@@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](O)[C@H]3O)[C@H](O[C@H]4CC[C@]5(C)[C@H]6CC[C@@]7(C)[C@@H](C[C@@H]8O[C@](O)(CCCCO[C@@H]9O[C@H](CO)[C@@H](O)[C@H](O)[C@H]9O)[C@@H](C)[C@H]78)[C@@H]6CC=C5C4)O[C@@H]2CO)[C@H](O)[C@H](O)[C@H]1O MHKGPHKABOLURA-JNVLQWCMSA-N 0.000 description 3
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000002924 anti-infective effect Effects 0.000 description 2
- 239000006286 aqueous extract Substances 0.000 description 2
- XIURVHNZVLADCM-IUODEOHRSA-N cefalotin Chemical compound N([C@H]1[C@@H]2N(C1=O)C(=C(CS2)COC(=O)C)C(O)=O)C(=O)CC1=CC=CS1 XIURVHNZVLADCM-IUODEOHRSA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 241001132374 Asta Species 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 210000001132 alveolar macrophage Anatomy 0.000 description 1
- 230000002509 aphrodisiac effect Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 244000039328 opportunistic pathogen Species 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
Classifications
-
- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Immunology (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Steroid Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a standardised steroid saponin mixture obtained from the epigeous part of Tribulus terrestris L., containing more than 80 per cent of furostanol saponins, a method of obtainment and its application as an immunostimulator and immunomodulator.
Description
LV13366
STANDARDISED STEROID SAPONIN MIXTURE, METHOD OF ITS OBTAINING AND APPLICATION
FIELD OF THE INVENTION
The present invention relates to a standardised steroid saponin mixture obtained from the epigeous part of Tribulus terrestris L for application in the synthesis of drugs exerting the already known aphrodisiac effect in addition to drugs with new indications. The invention pertains also to the method of preparing of a standardised steroid saponin mixture obtained from the epigeous part of Tribulus terrestris L
BACKGROUND OF THE INVENTION A method of isolating a standardised steroid saponin mixture obtained from the epigeous part of Tribulus terrestris L. (BG Patent 52085) is know to exist, the said method consisting in subjecting the plant material to extraction using an up to 70 per cent solution of low alcohol, ethanol in particular, then chloroform treatment and extraction from the vvater solution using vvater-saturated n-butanol, follovved by concentration of the butanol extract up to 1/8th of the volume, after vvhich the residue thus obtained is separated and dried, while the residues from the mother liquors are water-dissolved and then added directly to the chloroform-treated vvater solution. According to the examples described, a standardised mixture is thus obtained, the said mixture containing 50 to 55 per cent of furostanol saponins, calculated on the basis of protodioscin. Harvest is betvveen 1.7 to 2 per cent depending on the saponin content in the plant material.
The problem that the present invention has solved is the extraction from the epigeous part of Tribulus terrestris L. of a substance containing a high Ievel of furostanol saponins, vvhich can be used to prepare nevv drugs vvith nevv indications. 2
DETAILED DESCRIPTION OF THE INVENTION
According to the present invention, the problem has been solved by obtaining a standardised steroid saponin mixture containing 80 per cent of furostanol saponins. The method of preparation consists in subjecting the ground plant material to extraction using aqueous methanol solvent with a water/methanol ratio ranging from 1:1 to 1:99 at a temperature from 52 to 55°C. The extract is then treated with active carbon and after filtration is vacuum concentrated until methanol is fully removed. The aqueous concentrate is subsequently subjected to extraction with methylene chloride. Methylene chloride is then distilled off and regenerated for further use in the method. The purified aqueous solution is acidified with hydrochloric acid up to pH 4 and is subjected to extraction with vvater-saturated n-butanol by the known method. Butanol extracts are then vvashed with a sodium chloride solution and vacuum concentrated to obtain a dry residue vvhich is dissolved in a low alcohol. Thereafter, the concentrate is either added drop-by-drop to acetone and left at a temperature of 10-15° C for about 24 hours, then filtered, vvashed with acetone and dried, or vvashed with a 0.5-1.5-per cent sodium hydroxide solution and, after separating the two layers, the butanol layer is vvashed vvith vvater to reach pH = 7, then the butanol extracts are vacuum concentrated until the organic solvent is fully removed and the aqueous solution is dried. A soft and light bright yellow povvder containing more than 80 percent of furostanol saponins is thus obtained. Harvest from air-dried plant material is about 3 per cent.
This new product containing more than 80 per cent of furostanol saponins can be used to prepare new drugs exerting an immunomodulating and/or immunostimulating effect.
EXAMPLES EXAMPLE 1
One tonne of ground plant material is subjected to extraction using a 70- per cent methanol aqueous solution at 50° C until exhaustion. The extract is dravvn off, then active carbon is added thereto. The resultant is passed 3 3 LV 13366 through a filter press. The aqueous concentrate thus obtained is subjected to a triple extraction with methylene chloride in a ratio of 1:2, 1:1, and 1:1 respectively. The organic solvent is distilled of and regenerated, vvhile cube residue is disposed of. The purified aqueous extract is acidified with hydrochloric acid up to pH 4 and subjected to an eightfold extraction with water n-butanol in a ratio of 1:1. Butanol extracts are washed with a 5-per cent sodium chloride solution and the concentrated until a dry residue is obtained under vacuum at 60° C and then dissolved into methanol. The mixture is slowly dropped into acetone, left at a temperature of 10° C for 20 hours and finally Nutch-filtered, washed with acetone and dried. A light yellow powder containing 85 percent of furostanol saponins is obtained as a result. Harvest is 3 per cent from air-dried plant material.
The product analysis is performed on the basis of protodioscin. (R. Gyulemetova, M. Tomova, M. Simova, T. Pangarova, Pharmazie, 37, H.4 1982). EXAMPLE2
One tonne of ground plant material is subjected to extraction using a 70- per cent methanol aqueous solution at 50° C until exhaustion. The extract is drawn off, then active carbon is added thereto. The resultant is passed through a fiiter press. The aqueous concentrate thus obtained is subjected to a triple extraction with methylene chloride in a ratio of 1:2, 1:1, and 1:1 respectively. The organic solvent is distilled of and regenerated, vvhile cube residue is disposed of. The purified aqueous extract is acidified with hydrochloric acid up to pH 4 and subjected to an eightfold extraction with vvater saturated n-butanol in a ratio of 1:1. Butanol extracts are vvashed with a 1-per cent sodium hydroxide solution, the ratio of alkaline vvater to butanol extract being 1:1 and after separating the tvvo layers, the butanol layer is vvashed vvith vvater up to pH 7. Butanol extracts are then vacuum concentrated until the organic solvent is fully removed, after vvhich the aqueous solution is dried. A light yellow povvder containing 85 percent of furostanol saponins is obtained as a result. Harvest is 3 per cent from air-dried plant material. 4
The product analysis is performed on the basis of protodioscin. (R. Gyulemetova, M. Tomova, M. Simova, T. Pangarova, Pharmazie, 37, H.4 1982).
RESULTS FROM TESTING OF THE STEROID MIXTURE ACCORDING TO THE INVENTION ON THE IMMUNE SYSTEM OF EXPERIMENTAL ANIMALS
Material and method
Plant material
The substance containing furostanol saponins (FS) was isolated from Tribulus terrestris L (Zigophyllaceae) according to the present invention. Lyophilised FS were stored at 4°C away from light and moisture. The solution used the experiments was prepared ex tempore on the basis of physiologic solution.
Experimental animals and protocol
Male ICR mice (average body vveight from 18 to 20 g) were given various oral doses of FS, according to the description shown in Table 1.
Experimental infection
Experimental infection was caused in a mouse by subcutaneous inoculation of 25 to 30 bacterial celis from an 18-hour agar Kl. Pneumonae culture (strain No 52145, Pasteur Institute, Paris). Contamination dose was chosen after running preliminary experiments aimed at obtaining a 50-per cent survival rāte. The infection was followed up for eight days, taking account of the subject’s general condition, mortality rāte (percentage), survival rāte and average survival time (ASTa in days). The protective effect of FS was evaluated along the increase in survival rātes and average survival times calculated as differences betvveen experimental groups and Controls.
In-vitro antibacterial inhibition tēst
The sensitivity of Kl. Pneumoniae celi to FS was determined using an adapted version of the Bauer method (4). Petri dishes (100 mm) containing agar were inoculated with 0.2 ml of bacterial suspension (10 cells/ml). VVells (10 mm) were made and filled with 0.2 ml of a FS solution of various concentration. Inhibition areas were measured after incubation for 24 hours at 5 5 LV 13366 37°C. Keflin was used as a positive control (Lilly, USA), in minimum inhibition concentrations of 0.150 mg/ml.
Phagocytic and microbicidal activity of alveolar macrophages aMa
Alveolar Ma were obtained through in situ lung lavage by injecting and drawing a 0.1 -1.0 ml TCM 199 solution containing 20 mM HEPES, 100 U/ml of penicillin and 0.1 mg/ml of streptomycin (Flaw Lab., UK) plus 3 U/ml of heparin (G. Richter, Hungary). The average number of washed aMa from each group was determined and the celis were then placed on incubation plates (BDSL, Scottland) in a concentration of 3 x 10 cells/ml to assess their phagocytic and microbicidal activity using a H3-thymidine radiometric method.(5)
Results
Anti-infectious protective effect
The results thus obtained show that FS affect the course and outcome of an experimental Kl. Pneumonia-induced infection in mice. In treated animals, mortality rāte reached a maximum of 30 to 40 per cent by the 4^-5^ day from infection (Figurē 1). The peak in mortality rāte in the Controls was considerably higher (70 to 80 percent) and was observed on the 2nd - 3rd day from infection. To summarise, the administration of FS leads to reduced severity of the clinical picture, lower mortality rāte, and prolonged lifetime in treated animals.
The anti-infectious effect of FS variēs according to dosage and administration schedules (Table 1). The highest dose administrated (625.0 mg/kg) shovved the strongest protective effect in ali 10-day administration schedules, starting 35, 25 or 15 day before infection; the peak of the effect was observed where the last FS dose was administered on the 15th day before infection.
In contrast to the marked in-vivo activity, FS do not exhibit in-vitro antibacterial activity. A solution of FS in physiological serum at concentrations of 0.01, 0.025, 0.05, 0.1, 2.0, 5.0, 10.0, 25.0, 50.0, 100.0, 250.0 and 500.0 mg/ml did not suppress Kl. Pneumoniae grovvth, unlike the marked suppression obsen/ed in the Keflin group, within a mean area of 21.5 ± 1.0 mm. 6
Stimulation ofaMa
Doses of 625.0 mg/kg cause aMa to increase in number and phagocytic and microbicidal activity with a peak by the 20* day from the last application of FS (Table 2).
Discussion
Unlike other saponins (3), FS do not exert a bactericide effect. FS influence the main effector mechanisms of celi and humoral immunity thus granting a marked protection from infection. The optimum dose of 625.0 mg/kg administered over a sequence of 10 days is very close to the schedule (6) applied in humāns. The strongest protective effect obtained with the schedule vvhere the time gap betvveen the last application of FS and infection is 15 days can be explained with the dynamics of aMa activation. The maximum increase in their functional activity is observed by the 20* day vvhich coincides with the acute phase of the infection (day 3 to 5). The great number of aMa with an increased phagocytic and microbicidal capacity prevent aggravation of the infection and reduce mortality rāte, thus simultaneously increasing survival rāte. Obviously, the protective effect of FS is fully achieved only after a given period of time necessary for the immune system to react. FS activate aMa vvhich play a significant role in the anti-bacterial protection of the lung, in particular in managing infections caused by opportunistic pathogens such as Kl. Pneumonae (6).
References: 1. Yamada, H. (1991) Curr. Opin. Biotechnol., 2, 203. 2. VVagner, H. (1990) Pūre Appl. Chem. 131 q 117. 3. Cambell, J.B., Bede J.C. (1989) E.A.G. Lett., 5, 12. 4. Bauer, A.W., Kirby, W.M.M., Sherris, J.C., Truck, M. (1966) Am. J. Clin. 5. Dimov, V., Ivanovska, N., Bankova, V.V, Nikolov N, Popov, S. (1991) Apidologie 22,155. 6. Zarkova, S. (1984) Rev.Port. Cienc. Veter., 79,117. N- C0 o (Λ Φ Έ Φ Q O i_ Q σ c "»3 JO 2 E t» o c 2εε n c o o £ c ts c Φ E w_ Φ o. X Φ c o c CO Ό Φ O 2 Ό _C c/b φ cn ’c o E 2 Φ C 0. 2 o Φ cn 2 o o S c jo> .Ω <0 Protective effect <D ļ— CO < 0.8 co 6Ό 0.4 0.4 0.5 CM 3.5 0.4 t— esi 0.4 0.4 Iļb Μ ·γ Φ φ ? -s 1..=:(0 O 3 ία W 32.6 45.6 28.5 10.0 20.0 ’ 15.5 42.9 14.3 33.9 27.3 16.2 o o Total dosec 625.0 312.5 187.5 125.0 250.0 125.0 Treatment T reatmentD 15,14,13.12,11,10,9,8,7,6 25,24,23,24,21,20,19,18,17,16 35,34,33,32,31,30,29,28,27,26 10,9,8,7,6 20,19,18,17,16 30,29,28,27,26 I 18,17,16 9‘Z 17,16 I 27,26 25,23,21,19,17 25,24,23,22,21,20,19,18,17,16 π Φ cn O O 62.5 O'OS 12.5
co Ό O) "o) E, φ cn 0 Ό co LL _ Φ ra « Φ 25 >* cn 1 (0 O WQQ ra J3 O
O) -X ~ch E cn >ī co 2, φ £ Φ c5 ω > £ 2 «3 > 'ā 2 cn Φ O) co i— Φ >
cn co c c < < Φ 0) o> cn c c co co -c jC O O
The above results are typical data obtained from three independent experiments using different experimental animals. LV 13366 LV13366
Immunomodulating properties of FS
Table 2: Activation of aMa in FS-treated mice
Days of investigation a Number0 Phagocytosisc Microbicidal effectd 1 202± 62e 22.1 ±2.0® 10.1 ±1.2 5 213+ 81 e 25.0 ± 3.7 13.3 ±1.9 10 318±120 29.2 ±4.4 15.0 ±2.7 15 323 ± 202 33.0 ± 4.4 16.6 ±4.8 20 476± 88 37.2 ±1.7 18.0 ±4.7 30 371 ±191 32.2 ± 5.8 17.2 ±2.2 Controls r 176± 31 21.2 ±1.9 9.0 ±1.2 a Days from the last application of FS for 10 consecutive days in a doses of 62.5 mg/kg orally b In thousands (χ 103) c Percentage of aMa-phagocyted bacterial celis d Percentage of bacterial celis killed one hour after phagocytosis by aMa 6 Statistically unreliable (p> 0.05 by the Student tēst) f Controls were fed with physiological serum alone.
The results represent average data from four independent experiments using 10 experimental animals for each group.
Claims (3)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BG107499A BG65737B1 (en) | 2003-01-24 | 2003-01-24 | Standardized mixture of steroid saponins, method for the preparation and apllication thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| LV13366B true LV13366B (en) | 2006-03-20 |
Family
ID=32739195
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| LVP-05-96A LV13366B (en) | 2003-01-24 | 2005-08-16 | Standardised steroid saponin mixture, a method of its obtaining and application |
Country Status (10)
| Country | Link |
|---|---|
| AU (1) | AU2003286011A1 (en) |
| BG (1) | BG65737B1 (en) |
| EA (1) | EA008763B1 (en) |
| EE (1) | EE05443B1 (en) |
| GB (1) | GB2412867B (en) |
| HR (1) | HRPK20050614B3 (en) |
| LV (1) | LV13366B (en) |
| PL (1) | PL377142A1 (en) |
| UA (1) | UA80182C2 (en) |
| WO (1) | WO2004064852A1 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080182801A1 (en) | 2003-12-22 | 2008-07-31 | Btg International Limited | Core 2 glcnac-t inhibitors |
| GB0513881D0 (en) | 2005-07-06 | 2005-08-10 | Btg Int Ltd | Core 2 GLCNAC-T Inhibitors III |
| GB0329667D0 (en) | 2003-12-22 | 2004-01-28 | King S College London | Core 2 GlcNAc-T inhibitor |
| GB0513888D0 (en) | 2005-07-06 | 2005-08-10 | Btg Int Ltd | Core 2 GLCNAC-T Inhibitors II |
| CN103040880B (en) * | 2012-12-13 | 2017-10-10 | 大兴安岭林格贝寒带生物科技股份有限公司 | A kind of isolation and purification method of tribuloside |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BG52085B2 (en) * | 1985-01-18 | 1996-06-28 | Tomova | Method for the isolation of standardized mixture of steroid saponins |
| US6343258B1 (en) * | 1999-08-13 | 2002-01-29 | ALEXIS Brian | Method for testing for readiness for harvesting of tribulus terrestris l. having high steroidal saponin content |
-
2003
- 2003-01-24 BG BG107499A patent/BG65737B1/en unknown
- 2003-11-12 UA UAA200508283A patent/UA80182C2/en unknown
- 2003-12-11 AU AU2003286011A patent/AU2003286011A1/en not_active Abandoned
- 2003-12-11 PL PL377142A patent/PL377142A1/en unknown
- 2003-12-11 EE EEP200500024A patent/EE05443B1/en unknown
- 2003-12-11 GB GB0513893A patent/GB2412867B/en not_active Expired - Fee Related
- 2003-12-11 WO PCT/BG2003/000043 patent/WO2004064852A1/en not_active Application Discontinuation
- 2003-12-11 EA EA200501170A patent/EA008763B1/en unknown
-
2005
- 2005-07-01 HR HR20050614A patent/HRPK20050614B3/en not_active IP Right Cessation
- 2005-08-16 LV LVP-05-96A patent/LV13366B/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| GB2412867B (en) | 2007-07-04 |
| UA80182C2 (en) | 2007-08-27 |
| EE05443B1 (en) | 2011-08-15 |
| GB0513893D0 (en) | 2005-08-10 |
| BG107499A (en) | 2004-08-31 |
| HRP20050614A2 (en) | 2006-06-30 |
| AU2003286011A1 (en) | 2004-08-13 |
| WO2004064852A1 (en) | 2004-08-05 |
| BG65737B1 (en) | 2009-09-30 |
| EA008763B1 (en) | 2007-08-31 |
| WO2004064852B1 (en) | 2004-10-07 |
| HRPK20050614B3 (en) | 2007-10-31 |
| EE200500024A (en) | 2005-10-17 |
| PL377142A1 (en) | 2006-01-23 |
| EA200501170A1 (en) | 2005-12-29 |
| GB2412867A (en) | 2005-10-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sajeevan et al. | Dose/frequency: a critical factor in the administration of glucan as immunostimulant to Indian white shrimp Fenneropenaeus indicus | |
| KR100900725B1 (en) | A preparation method of artemisia extract containing high content of eupatilin | |
| LV13366B (en) | Standardised steroid saponin mixture, a method of its obtaining and application | |
| Yadav et al. | Anthelmintic and Antibacterial Activity of Psidium Guajava Leaf Extracts | |
| CN110724121B (en) | Bibenzyl derivative in dendrobium officinale leaf and preparation method and application thereof | |
| KR101394550B1 (en) | Anti-bacterial or Anti-inflammatory Composition Comprising Extracts from Flower of Rosa hybrida as Active Ingredient | |
| WO1997004791A1 (en) | Antiviral agent and process for preparing the same | |
| KR20150129344A (en) | Anti-obese and anti-diabetic composition comprising fingerroot polysaccharides | |
| CN108938724B (en) | Neem leaf extract and preparation process and application thereof | |
| KR101703352B1 (en) | Artemisia extract | |
| RU2385159C2 (en) | Method for making preparation of jagel-m with antituberculous action | |
| WO2003028746A1 (en) | Method for producing extracts of pelargonium sidoides and/or pelargonium reniforme, and the use of said extracts | |
| CN111840340A (en) | Application of pomegranate bark tannin in preparation of medicine for inhibiting bacterial quorum sensing system | |
| CN113521262A (en) | Lysozyme preparation with anti-inflammatory effect | |
| JPH08151329A (en) | Antiviral agent, carcinostatic agent and production thereof | |
| KR102094228B1 (en) | Composition for preventing or treating hearing loss comprising Uncaria rhynchophylla extracts | |
| CN103432214A (en) | Preparation method and application of effective components of polygonum capitatum | |
| KR20150129343A (en) | Anti-obese composition comprising Java tumeric polysaccharides | |
| KR102131044B1 (en) | Composition for preventing or treating hearing loss | |
| KR102313249B1 (en) | Composition for preventing or treating hearing loss comprising extract or fraction of Castanopsis echinocarpa | |
| LT5340B (en) | Standardised steroid saponin mixture, a method of its obtaining and application | |
| CN110172016B (en) | Benzyl alcohol compound, pharmaceutical composition and application thereof | |
| KR102568412B1 (en) | Method for producing of milk thistle extract with increased silymarin bioavailability | |
| RU2710270C1 (en) | Agent having immunomodulatory activity, and a method for production thereof | |
| WO2024096280A1 (en) | Composition for preventing, ameliorating, or treating alzheimer's disease containing as active ingredient vitis vinifera stem extract or compound isolated therefrom |