CA2321783A1 - Use of an expressed juice, digestive juice or extract from flesh-eating plants for the inhibition of protein kinases - Google Patents
Use of an expressed juice, digestive juice or extract from flesh-eating plants for the inhibition of protein kinases Download PDFInfo
- Publication number
- CA2321783A1 CA2321783A1 CA002321783A CA2321783A CA2321783A1 CA 2321783 A1 CA2321783 A1 CA 2321783A1 CA 002321783 A CA002321783 A CA 002321783A CA 2321783 A CA2321783 A CA 2321783A CA 2321783 A1 CA2321783 A1 CA 2321783A1
- Authority
- CA
- Canada
- Prior art keywords
- juice
- flesh
- extract
- plants
- eating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
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
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the use of a press-juice, digestive juice or extract of carnivorous plants for the production of a medicine for inhibiting protein kinases.
Description
Use of aQ expressed juice, digestive juice or extract from flesh-eating plants for the inhibition of protein kinases The invention relates to the inhibition of protein kinases through the use of an expressed juice, digestive juice or extract from flesh-eating plants Recent research in the field of drug active substances has been concentrated on the field of drugs produced on a natural basis. Besides the surprisingly great variety of active substances that can be found in plants and naturally occurring microorganisms, drugs produced on a natural basis often have the surprising advantage of better tolerance compared to synthetically produced active substances, which often provoke severe, undesired side-effects. The flesh-eating plants are an interesting example of a source for natural active substances. Thus for example the expressed juice of certain flesh-eating plants is used for the control of malignant diseases such as cancer, Crohn's disease, ulcerative colitis, herpes and aids.
This is for example stated in EP-A-249465, EP-A-219295 and DE-A-2920631.
At the same time, there is a whole range of diseases in which the conventional medication with synthetic active substances shows practically no result. Gastric ulcers are an example of such a disease, for the control of which costly chemotherapeutic treatments are now given. The therapies currently most commonly used at present are the modified triplet therapies. These therapies are for example described in the technical journal "hunch. med.
WSCHR., 140 (1998), No.7, pp 90 to 93" and in "Deutsches Arztemagazin No.51/52, December 1997, der Kassenarzt" on pages 31 to 34. These therapies have however the disadvantage that, like comparable chemotherapies in cancerous diseases, they are accompanied with severe side-effects.
Hence the purpose of the present invention is to provide an active substance or an active substance combination with which it is possible to control diseases which respond to the inhibition of protein kinases, with as far as possible no occun-ence of side-effects.
This is for example stated in EP-A-249465, EP-A-219295 and DE-A-2920631.
At the same time, there is a whole range of diseases in which the conventional medication with synthetic active substances shows practically no result. Gastric ulcers are an example of such a disease, for the control of which costly chemotherapeutic treatments are now given. The therapies currently most commonly used at present are the modified triplet therapies. These therapies are for example described in the technical journal "hunch. med.
WSCHR., 140 (1998), No.7, pp 90 to 93" and in "Deutsches Arztemagazin No.51/52, December 1997, der Kassenarzt" on pages 31 to 34. These therapies have however the disadvantage that, like comparable chemotherapies in cancerous diseases, they are accompanied with severe side-effects.
Hence the purpose of the present invention is to provide an active substance or an active substance combination with which it is possible to control diseases which respond to the inhibition of protein kinases, with as far as possible no occun-ence of side-effects.
This purpose is achieved through the use of an expressed juice, digestive juice or extract of flesh-eating plants for the inhibition of protein kinases. These are also referred to below collectively as juices.
Protein kinases play a central role in the growth, division and differentiation of cells. Growth signals, which in the form of growth factors bind to specific recognition structures (receptors) on the outer side of the cell membrane, are transmitted into the cell nucleus via various kinase cascades, and there lead to the modification of transcription signals. As a consequence of this, specific genes become activated or the transcription of other genes is stopped.
Surprisingly, it has now been found that the expressed juice, an extract or the digestive juice from flesh-eating plants makes inhibition of protein kinases possible, simultaneously making it possible to control various diseases, previously treatable only with very great difficulty.
In particular, it has been found that the juices of flesh-eating plants show effects which resemble those of conventional antibiotics, but with the occurrence of markedly fewer side-effects. The juices of flesh-eating plants show action against gram-positive and gram-negative bacteria, fungi and other germs.
In particular, the juices used according to the invention are suitable for the control of plasmodia, chlamydia, trypansomes, phototropic bacteria, staphylococci, streptococci, pneumococci, klebsiella, candida, mucor, aspergillus and bacterium pyocyaneus.
Presumably the aforesaid germs, bacteria and fungi are inactivated by inhibition of the protein kinases. In particular, the expressed juice, an extract or the digestive juice of flesh-eating plants are suitable for the control of helicobacter pylori, one of the conunonest causes of gastric ulcers.
By series of experiments, it was further confirmed that inhibition of the following germs, fungi and bacteria can be achieved by means of the juice of flesh-eating plants:
Pseudomonas aeruginosa, Proteus mirabilis, Staphylococcus aureus, Streptococcus pneum-oniae, Escherichia coli, Staphylococcus epidermis, group B Streptococci (haemolysing), Streptococcus pyogenes (A), Candida albicans, Aspergillus fumigatus, Pneumococcus mucosus, Bact. pyocyaneus, Bact. Proteus vulgaris, Klebsiella oraeanae, Candida mycod., Candida tropicaus and Mucor racemosus.
The obtention of an expressed juice, digestive juice or extract of flesh-eating plants, and also preferred presentations, are discussed below.
The flesh-eating plant can be any flesh-eating plant. Preferably, it belongs one of the genera Heliamphora, Sarracenia, Darlingtonia, Cephalotus or Nephentes. Also preferred are the flesh-eating plants of the Drosera family, especially Dionaea muscipula, the Venus flytrap.
The expressed juice, an extract or the digestive juice of flesh-eating plants can be used.
The use of the digestive juice has the advantage that the plants do not have to be destroyed for its obtention, since the digestive juice can be tapped off. Here a significant increase in the yield can be achieved by constant feeding of the plants. The flesh-eating plants are usually fed with meat fragments of the same tissue over a long period. This also has the result that the composition of the digestive juice is stabilised, i. e. that the individual components of the digestive juice in the same type of plants are present in similar proportions.
Without feeding, this is not the case, so that the active substances are present in different concentrations from plant to plant, and hence the therapeutic effect only leads to a therapeutic result by mixing the digestive juices of several plants. In order to increase the quantity of digestive secretions which are supplied by the plants, a stimulation of the secretion can be performed. For this the spraying of the plants with uric acid has been found to be particularly suitable, the remaining residues of uric acid showing no adverse effects. Further suitable agents for stimulation of the secretion are bactopeptones, kalifora, glutamic acid, yeast extract, peptones, taurine, proline and asparagine. The removal of the digestive juice is conveniently effected by pipetting off or by making small puncture wounds on the inner side of folds, and subsequent collection of the secretion in micropipettes.
However, according to the invention the expressed juice can also be used. The expressed juice is advantageously produced using fresh plants, i.e. plants in which no enzymatically triggered wilting process has yet begun. Usually, the harvested plants are first washed, which removes extraneous components. The cleaned plants are then chopped, grated or mashed, and then pressed in the wet state. Advantageously, they can be treated for a few seconds with pressurised steam before pressing, in order to inactivate enzymes, effect a disintegration by partial destruction of the cell walls and, through albumin precipitation, to facilitate a later clarification process. Similariy, shock freezing can be performed.
For the pressing, both continuously operating fly-presses, hydraulic filter presses and also continuous screw presses are commonly used. The parameters to be set for this and other process operations are well known to the specialist. The expressed juice can then be treated by normal processes, such as filtration, centrifugation and separation. If necessary, it is next ultrapasteurised and then filled into sterile bottles. Next it is preferably cooled as quickly as possible to temperatures below 70°C.
Also usable are extracts from flesh-eating plants. Here, the following extractants and processes can be used:
Eatractants:
cold water, hot water, dilute acetic acid, sweet wine, ethanol-water mixtures, ethanol and fatty oils or neutral oils and supercritical gases, for example supercritical CO2.
Extraction Processes:
static maceration, stirred maceration, turbulent extraction, digestion, percolation, repercolation, evacolation, diacolation, combinations of maceration and percolation, ultrasonic extraction, countercurrent extraction and extraction with separators, centrifuges and decanters.
The equipment to be used for this and the process parameters normally used are familiar to the specialist.
The products obtained by tapping off the digestive juice, by pressing and by extraction can be further processed in normal ways. In particular, these juices can be freeze-dried, especially lyophilised. For this, the usual additives, such as mannitol or glycerol can be used during the lyophilisation. The equipment needed for this and the process parameters are well known to the specialist. Further, various additives which have a favourable influence on the therapeutic action on administration, such as for example formic acid, benzoic acid, citric acid, malic acid, cyanidine-3-glucoside, sodium, magnesium, potassium, calcium, chlorine, naphthoquin-ones, L-arginine, L-asparagine, L-serine, L-glutamic acid, L-alanine, L-threonine, L-lysine, glycine, L-tyrosine, L-phenylalanine, L-valine, L-leucine and L-isoleucine, can be added to these juices.
The juices of flesh-eating plants can be administered in various ways.
The juices can be used directly orally or topically; alternatively, they can previously be diluted with a harmless solvent. Examples of suitable solvents are water, sweet wine, ethanol-water mixtures and salt solutions.
Usually, they are diluted to a dry residue of 0.1 to 10 wt %, preferably 0.5 to 5 wt %, and especially preferably 0.8 to 2.5 wt %.
Further, the juice of the flesh-eating plant can be administered orally in the form of tablets and capsules. The equipment needed for this and the processes for formulation are well known to the specialist. Further, topical applications in the form of the juices themselves or in the form of ointments are also possible. The equipment and the materials and processes needed for the production of ointments are well known to the specialist.
Further, it is also possible to produce parenteral formulations from the juices ofthe flesh-eating drugs. These can be administered intramuscularly, subcutaneously or intravenously.
The equipment, additives and processes needed for the production of such parenteral formul-ations are well known to the specialist.
The following experiments illustrate the invention:
1. Inhibition of Protein Kinases Sample material Carnivora expressed juice dilute and also C:arnivora~
expressed juice neat, produced by Carnivora Research GmbH.
Until the start of the experiment, both samples were stored unopened in the refrigerator. On the day of the first experiment, both samples were each prediluted 1:10 and 1:100 with water and DMSO, so that a 1:10 and a 1:100 initial solution of both samples in 10 %
DMSO was to hand. On addition to the test, a further 1:10 dilution step was performed on each occasion.
Both samples were tested at 1:100 and 1:1000 dilutions. The DMSO concentration in the assay was 1 %. The prediluted initial solutions were kept at 4°C and used for all measure-ments over a period of 10 days.
Protein Kinases Used:
Cyclin-dependent kinase 4 (CDK 4) and cyclin-dependent kinase 2 (CDK 2) Tyrosine kinase of the (i receptor of platelet-derived growth factor (PDGF-R(3-TK) Tyrosine kinase of the receptor for epidermal growth factor (EGF-R-TK) Tyrosine kinase of the ERBB2 receptor ERBB2-TK
Tyrosine kinase of the receptor-1 of fibroblast growth factar (FGF-R1-TK) Tyrosine kinase of insulin-like growth factor receptor, type 1 (IGF-1-R-TK) Tyrosine kinase of the insulin receptor Ins-R-TK
Protein kinase C, subtype y (PKC-y) Protein kinase C, subtype E (PKC-e) Janus kinase 2 (JAK 2).
The protein kinases were cloned from the RNA of suitable cells by means of RT-PCR. The isolated DNA sections were then subcloned in baculovirus transfer vectors and recombinant baculoviruses were then produced. Using the recombinant baculoviruses, the proteins were expressed in Sf~ insect cells. The kinases were purified from the insect cell lysates using glutathione agarose gel.
All kinase assays were performed in 96-well microtitre plates in a reaction volume of 100 ~l.
For the measurement of PKC activity, the test (mixture] contained 50 mM HEPES-NaOH, pH
7.5, 1 mM EDTA, 1.25 mM EGTA, 5 mM MgCl2, 1.32 rr>Nf CaCl2, 5 pg/ml phosphatidyl-serine, 1 ~g/ml 1,2-diolein, 1 mM DTT, 0.1 uM [33P]-ATP, 50 ng recombinant kinase and 0.5 ~g histone Hl as substrate. For all other kinases, the test mixture contained 50 mM
HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnClz, 3 u.M Na orthovanadate, 1 mM DTT, 0.1 I,WI ATP, 100 ng recombinant kinase and a suitable substrate.
The reaction mixture was incubated for 80 mins at 30°C. The reaction was stopped by addition of 100 ul H3P0~ (2 %). Next the plates were washed three times with HZO. The incorporation of [33P)-phosphate into the relevant substrate was determined using a scintill-ation counter for microtitre plates. The results are summarised in the following table.
Table 1 Percentage Action (Inhibition) Kinase dilute expressed juice neat expressed juice 1:100 1:100 1:1000 CDK4/cyclin D - 66 % - 109 % - 41 CDK2/cyclin E _ gg %
PDGF-R(3-TK -78 % - 98 % - 64 EGF-R-TK -78 % - 96 % - 68 ERbB2-TK - 62 % - 88 % - 82 FGF-RI-TK - 70 % - 100 % - 67 IGF-1-R-TK - 59 % - 9Ei % - 66 Ins-R-TK - 68 % - 99 % - 7~
PKC-y -57% _34%
'' PKC-E - 6~ % -. _ 37 J~ ~ -81% ~% -79%
The results clearly show that both the neat expressed juice from Carnivora and also a diluted expressed juice cause inhibition of protein kinases.
II. Control of Helicobacter pylori Studies relating to the eradication of HeIicobacter pylori were carried out on the duodenal ulcer model in the rat. In this ulcer model, rats which are predominantly colonised with Helicobacter pylori were used. After oral administration of t:he ulcerogen cysteamine hydrochloride, duodenal ulcers and gastric ulcers were 100 °,%
detectable macroscopically and microscopically 48 hrs after administration.
Carnivora~' expressed juice (2 % dry residue) was administered orally in the 3 dosages 0.06, 0.6 and 2.83 ml/kg body weight (BW) 3 x daily over a period of 20 days after ulcer induction.
The same administration regime was used for the reference group (combination of tetracycline (6 mg/kg BW), metronidazole (15 mg/kg BW) and basic bismuth nitrate (30 mg/kg BW)) and for the controls (isotonic common salt solution, 0.9 %
NaCI).
2, 5, 8, 14 and 21 days after the ulcer induction, the respective groups were examined for ulcer healing by macroscopic assessment, and in addition a determination of Helicobacter pylori was performed.
The following parameters were covered:
mean assessment score (mAS) based on the degree of severity of the duodenal and gastric ulcers, percentage frequency of the ulcers (% U), ulcer index (UI) (mAS x %
U), area of the ulcers, body weight and mortality.
In this model, the combination of the three reference substances showed the expected action.
The ulcer index was lowered, the lesion areas of the duodenal or gastric ulcers were decreased and the incidence of Helicobacter pylori was lowered.
The three dosages of Carnivora expressed juice tested showed very good dose-dependent effects. The ulcer index was lowered, and the lesion areas of the duodenal or gastric ulcers were decreased, or the ulcers completely eliminated. The incidence of Helicobacter pylori could be lowered, and at the highest dosage the eradication of this germ was effected. Further beneficial effects of the test substance manifested themselves in a decrease in the spontaneous deaths occurring during the first 3 days after administration of the ulcerogen and a greater weight gain compared to the controls over the test period from 0-21 days.
The demonstrated effects of Carnivora expressed juice confirm its outstanding action on this model.
Table 2 Group 2 5 8 14 21 days post ulcerogen DU GU DU GU DU GU DU GU DU GU
controlmAS 2.2 1.9 2.6 18 2.7 1.6 2.2 0.6 2.1 1.2 sd 0.-10.5 0.4 0.9 0.5 1.5 0.3 0.8 0.4 0.7 UI 219.0191.0258.0 204.0268.096.0 220.024.8 206.096.0 referencemAS -- -- 2.1 2.0 1.3 l.3 0.3 0.2 0.5 0.3 sd -- -- 0.5 0.5 1.3 1.3 0.7 0.5 0.8 0.6 UI -- -- 206.0 204.076.8 76.8 6.0* 4.4 21.6*5.2 dose m.~.S-- -- 2.:1 1.8 2.2 1:.3 1.8 0.8 1.4 0.6 sd -- -- 0.6 0.9 0.5 1..2 1.1 1.3 0.9 0.9 U -- -- 100 100 100 ti0 80 40 80 40 UI -- -- 238.0 178.0218.078.0 145.633.6 115.225.6 dose mAS -- -- 2.1 1.6 1.1 1.1 0.6 0.7 1.2 1.0 sd -- -- 0.7 1.1 1.0 I.0 0.8 0.9 1.1 1.0 U -- -- 100 80 60 fi0 40 40 60 60 UI -- -- 214.0 131.267.2*6:3.624.8*26.4 69.6 62.4 dose mAS -- -- 2.1 1.3 1.2 0.9 0.7 0.9 1.1 0.7 sd -- -- 0.2 0.9 1.2 0.9 1.0 0.9 1.0 0.7 UI -- -- 208.0*107.274.4*5fi.428.8*55.2 66.0 43.2 AS assessment score mAS mean assessment score U percentage of animals with ulcers of all degrees of severity UI ulcer index significance calculation, t-test, p<0.05, versus control Table 3 Area of duodenal ulcer [mmZ]
days post 2 5 8 1.1 21 ulcerogen Group r/n A r/n A r/n A r/n A r/n A
Control 8110 .10.95/5 25.1 4/5 17.5 4/5 13.0 315 14.9 Reference -- -- 3/5 20.9 2/5 21.2 0/5 0 O/S 0 Dose 1 -- -- 4/5 23.4 4/5 12.6 3/5 11.3 2/5 5.9 Dose 2 -- -- 3/5 39.8 2/5 4.7 0/5 0 1/5 3.1 Dose 3 -- -- 4/5 5.7 2/5 8.3 1/5 3.1 0/5 0 Table 4 Area of gastric ulcer [mm2]
days post 2 5 8 14 21 ulcerogen Group r/n A r/n A r/n A r/n A r/n A
Control 5/10 207.33/5 130.93/5 110.00/5 0 1/6 78.5 Reference -- -- 4/5 90.4 2/5 53.4 O/S 0 O/S 0 Dose 1 -- -- 3/5 238.83/5 93.2 1/5 113.11/5 28.3 Dose 2 -- -- 215 64.4 1/5 28.3 0/5 0 115 12.6 Dose 3 -- -- 2/5 28.3 1/5 12.6 0/5 0 O/S 0 r/n number of animals with ulcers of severity degree 2-3/number of animals studied A area (determined for ulcers of severity degree 2-3).
Table 5 _ . Helicobacter pylori (Hp+) [%]
days post 2 5 8 14 21 ulcerogen Group Control 80 100 100 80 80 Reference -- 60 40 20 20 Dose 1 -- 100 60 60 60 Dose 2 -- 60 :l0 40 40 Dose 3 -- 60 20* 0* 0*
* significance calculation, exact Fisher Test, p < 0.05, one-sided, versus control Table 6 Mortality days post 1 2 3 Total uicerogen e/n e/n e/n e/n Group Control 1/30 3/30 x/30 9130 Reference 1/20 0/20 0/20 1/20 Dose 1 0/20 1/20 1/20 2120 Dose 2 1/20 0120 0/20 1120 Dose 3 0/20 0/20 0/20 0/20 e/n number of animals died/total number of animals per group
Protein kinases play a central role in the growth, division and differentiation of cells. Growth signals, which in the form of growth factors bind to specific recognition structures (receptors) on the outer side of the cell membrane, are transmitted into the cell nucleus via various kinase cascades, and there lead to the modification of transcription signals. As a consequence of this, specific genes become activated or the transcription of other genes is stopped.
Surprisingly, it has now been found that the expressed juice, an extract or the digestive juice from flesh-eating plants makes inhibition of protein kinases possible, simultaneously making it possible to control various diseases, previously treatable only with very great difficulty.
In particular, it has been found that the juices of flesh-eating plants show effects which resemble those of conventional antibiotics, but with the occurrence of markedly fewer side-effects. The juices of flesh-eating plants show action against gram-positive and gram-negative bacteria, fungi and other germs.
In particular, the juices used according to the invention are suitable for the control of plasmodia, chlamydia, trypansomes, phototropic bacteria, staphylococci, streptococci, pneumococci, klebsiella, candida, mucor, aspergillus and bacterium pyocyaneus.
Presumably the aforesaid germs, bacteria and fungi are inactivated by inhibition of the protein kinases. In particular, the expressed juice, an extract or the digestive juice of flesh-eating plants are suitable for the control of helicobacter pylori, one of the conunonest causes of gastric ulcers.
By series of experiments, it was further confirmed that inhibition of the following germs, fungi and bacteria can be achieved by means of the juice of flesh-eating plants:
Pseudomonas aeruginosa, Proteus mirabilis, Staphylococcus aureus, Streptococcus pneum-oniae, Escherichia coli, Staphylococcus epidermis, group B Streptococci (haemolysing), Streptococcus pyogenes (A), Candida albicans, Aspergillus fumigatus, Pneumococcus mucosus, Bact. pyocyaneus, Bact. Proteus vulgaris, Klebsiella oraeanae, Candida mycod., Candida tropicaus and Mucor racemosus.
The obtention of an expressed juice, digestive juice or extract of flesh-eating plants, and also preferred presentations, are discussed below.
The flesh-eating plant can be any flesh-eating plant. Preferably, it belongs one of the genera Heliamphora, Sarracenia, Darlingtonia, Cephalotus or Nephentes. Also preferred are the flesh-eating plants of the Drosera family, especially Dionaea muscipula, the Venus flytrap.
The expressed juice, an extract or the digestive juice of flesh-eating plants can be used.
The use of the digestive juice has the advantage that the plants do not have to be destroyed for its obtention, since the digestive juice can be tapped off. Here a significant increase in the yield can be achieved by constant feeding of the plants. The flesh-eating plants are usually fed with meat fragments of the same tissue over a long period. This also has the result that the composition of the digestive juice is stabilised, i. e. that the individual components of the digestive juice in the same type of plants are present in similar proportions.
Without feeding, this is not the case, so that the active substances are present in different concentrations from plant to plant, and hence the therapeutic effect only leads to a therapeutic result by mixing the digestive juices of several plants. In order to increase the quantity of digestive secretions which are supplied by the plants, a stimulation of the secretion can be performed. For this the spraying of the plants with uric acid has been found to be particularly suitable, the remaining residues of uric acid showing no adverse effects. Further suitable agents for stimulation of the secretion are bactopeptones, kalifora, glutamic acid, yeast extract, peptones, taurine, proline and asparagine. The removal of the digestive juice is conveniently effected by pipetting off or by making small puncture wounds on the inner side of folds, and subsequent collection of the secretion in micropipettes.
However, according to the invention the expressed juice can also be used. The expressed juice is advantageously produced using fresh plants, i.e. plants in which no enzymatically triggered wilting process has yet begun. Usually, the harvested plants are first washed, which removes extraneous components. The cleaned plants are then chopped, grated or mashed, and then pressed in the wet state. Advantageously, they can be treated for a few seconds with pressurised steam before pressing, in order to inactivate enzymes, effect a disintegration by partial destruction of the cell walls and, through albumin precipitation, to facilitate a later clarification process. Similariy, shock freezing can be performed.
For the pressing, both continuously operating fly-presses, hydraulic filter presses and also continuous screw presses are commonly used. The parameters to be set for this and other process operations are well known to the specialist. The expressed juice can then be treated by normal processes, such as filtration, centrifugation and separation. If necessary, it is next ultrapasteurised and then filled into sterile bottles. Next it is preferably cooled as quickly as possible to temperatures below 70°C.
Also usable are extracts from flesh-eating plants. Here, the following extractants and processes can be used:
Eatractants:
cold water, hot water, dilute acetic acid, sweet wine, ethanol-water mixtures, ethanol and fatty oils or neutral oils and supercritical gases, for example supercritical CO2.
Extraction Processes:
static maceration, stirred maceration, turbulent extraction, digestion, percolation, repercolation, evacolation, diacolation, combinations of maceration and percolation, ultrasonic extraction, countercurrent extraction and extraction with separators, centrifuges and decanters.
The equipment to be used for this and the process parameters normally used are familiar to the specialist.
The products obtained by tapping off the digestive juice, by pressing and by extraction can be further processed in normal ways. In particular, these juices can be freeze-dried, especially lyophilised. For this, the usual additives, such as mannitol or glycerol can be used during the lyophilisation. The equipment needed for this and the process parameters are well known to the specialist. Further, various additives which have a favourable influence on the therapeutic action on administration, such as for example formic acid, benzoic acid, citric acid, malic acid, cyanidine-3-glucoside, sodium, magnesium, potassium, calcium, chlorine, naphthoquin-ones, L-arginine, L-asparagine, L-serine, L-glutamic acid, L-alanine, L-threonine, L-lysine, glycine, L-tyrosine, L-phenylalanine, L-valine, L-leucine and L-isoleucine, can be added to these juices.
The juices of flesh-eating plants can be administered in various ways.
The juices can be used directly orally or topically; alternatively, they can previously be diluted with a harmless solvent. Examples of suitable solvents are water, sweet wine, ethanol-water mixtures and salt solutions.
Usually, they are diluted to a dry residue of 0.1 to 10 wt %, preferably 0.5 to 5 wt %, and especially preferably 0.8 to 2.5 wt %.
Further, the juice of the flesh-eating plant can be administered orally in the form of tablets and capsules. The equipment needed for this and the processes for formulation are well known to the specialist. Further, topical applications in the form of the juices themselves or in the form of ointments are also possible. The equipment and the materials and processes needed for the production of ointments are well known to the specialist.
Further, it is also possible to produce parenteral formulations from the juices ofthe flesh-eating drugs. These can be administered intramuscularly, subcutaneously or intravenously.
The equipment, additives and processes needed for the production of such parenteral formul-ations are well known to the specialist.
The following experiments illustrate the invention:
1. Inhibition of Protein Kinases Sample material Carnivora expressed juice dilute and also C:arnivora~
expressed juice neat, produced by Carnivora Research GmbH.
Until the start of the experiment, both samples were stored unopened in the refrigerator. On the day of the first experiment, both samples were each prediluted 1:10 and 1:100 with water and DMSO, so that a 1:10 and a 1:100 initial solution of both samples in 10 %
DMSO was to hand. On addition to the test, a further 1:10 dilution step was performed on each occasion.
Both samples were tested at 1:100 and 1:1000 dilutions. The DMSO concentration in the assay was 1 %. The prediluted initial solutions were kept at 4°C and used for all measure-ments over a period of 10 days.
Protein Kinases Used:
Cyclin-dependent kinase 4 (CDK 4) and cyclin-dependent kinase 2 (CDK 2) Tyrosine kinase of the (i receptor of platelet-derived growth factor (PDGF-R(3-TK) Tyrosine kinase of the receptor for epidermal growth factor (EGF-R-TK) Tyrosine kinase of the ERBB2 receptor ERBB2-TK
Tyrosine kinase of the receptor-1 of fibroblast growth factar (FGF-R1-TK) Tyrosine kinase of insulin-like growth factor receptor, type 1 (IGF-1-R-TK) Tyrosine kinase of the insulin receptor Ins-R-TK
Protein kinase C, subtype y (PKC-y) Protein kinase C, subtype E (PKC-e) Janus kinase 2 (JAK 2).
The protein kinases were cloned from the RNA of suitable cells by means of RT-PCR. The isolated DNA sections were then subcloned in baculovirus transfer vectors and recombinant baculoviruses were then produced. Using the recombinant baculoviruses, the proteins were expressed in Sf~ insect cells. The kinases were purified from the insect cell lysates using glutathione agarose gel.
All kinase assays were performed in 96-well microtitre plates in a reaction volume of 100 ~l.
For the measurement of PKC activity, the test (mixture] contained 50 mM HEPES-NaOH, pH
7.5, 1 mM EDTA, 1.25 mM EGTA, 5 mM MgCl2, 1.32 rr>Nf CaCl2, 5 pg/ml phosphatidyl-serine, 1 ~g/ml 1,2-diolein, 1 mM DTT, 0.1 uM [33P]-ATP, 50 ng recombinant kinase and 0.5 ~g histone Hl as substrate. For all other kinases, the test mixture contained 50 mM
HEPES-NaOH, pH 7.5, 3 mM MgCl2, 3 mM MnClz, 3 u.M Na orthovanadate, 1 mM DTT, 0.1 I,WI ATP, 100 ng recombinant kinase and a suitable substrate.
The reaction mixture was incubated for 80 mins at 30°C. The reaction was stopped by addition of 100 ul H3P0~ (2 %). Next the plates were washed three times with HZO. The incorporation of [33P)-phosphate into the relevant substrate was determined using a scintill-ation counter for microtitre plates. The results are summarised in the following table.
Table 1 Percentage Action (Inhibition) Kinase dilute expressed juice neat expressed juice 1:100 1:100 1:1000 CDK4/cyclin D - 66 % - 109 % - 41 CDK2/cyclin E _ gg %
PDGF-R(3-TK -78 % - 98 % - 64 EGF-R-TK -78 % - 96 % - 68 ERbB2-TK - 62 % - 88 % - 82 FGF-RI-TK - 70 % - 100 % - 67 IGF-1-R-TK - 59 % - 9Ei % - 66 Ins-R-TK - 68 % - 99 % - 7~
PKC-y -57% _34%
'' PKC-E - 6~ % -. _ 37 J~ ~ -81% ~% -79%
The results clearly show that both the neat expressed juice from Carnivora and also a diluted expressed juice cause inhibition of protein kinases.
II. Control of Helicobacter pylori Studies relating to the eradication of HeIicobacter pylori were carried out on the duodenal ulcer model in the rat. In this ulcer model, rats which are predominantly colonised with Helicobacter pylori were used. After oral administration of t:he ulcerogen cysteamine hydrochloride, duodenal ulcers and gastric ulcers were 100 °,%
detectable macroscopically and microscopically 48 hrs after administration.
Carnivora~' expressed juice (2 % dry residue) was administered orally in the 3 dosages 0.06, 0.6 and 2.83 ml/kg body weight (BW) 3 x daily over a period of 20 days after ulcer induction.
The same administration regime was used for the reference group (combination of tetracycline (6 mg/kg BW), metronidazole (15 mg/kg BW) and basic bismuth nitrate (30 mg/kg BW)) and for the controls (isotonic common salt solution, 0.9 %
NaCI).
2, 5, 8, 14 and 21 days after the ulcer induction, the respective groups were examined for ulcer healing by macroscopic assessment, and in addition a determination of Helicobacter pylori was performed.
The following parameters were covered:
mean assessment score (mAS) based on the degree of severity of the duodenal and gastric ulcers, percentage frequency of the ulcers (% U), ulcer index (UI) (mAS x %
U), area of the ulcers, body weight and mortality.
In this model, the combination of the three reference substances showed the expected action.
The ulcer index was lowered, the lesion areas of the duodenal or gastric ulcers were decreased and the incidence of Helicobacter pylori was lowered.
The three dosages of Carnivora expressed juice tested showed very good dose-dependent effects. The ulcer index was lowered, and the lesion areas of the duodenal or gastric ulcers were decreased, or the ulcers completely eliminated. The incidence of Helicobacter pylori could be lowered, and at the highest dosage the eradication of this germ was effected. Further beneficial effects of the test substance manifested themselves in a decrease in the spontaneous deaths occurring during the first 3 days after administration of the ulcerogen and a greater weight gain compared to the controls over the test period from 0-21 days.
The demonstrated effects of Carnivora expressed juice confirm its outstanding action on this model.
Table 2 Group 2 5 8 14 21 days post ulcerogen DU GU DU GU DU GU DU GU DU GU
controlmAS 2.2 1.9 2.6 18 2.7 1.6 2.2 0.6 2.1 1.2 sd 0.-10.5 0.4 0.9 0.5 1.5 0.3 0.8 0.4 0.7 UI 219.0191.0258.0 204.0268.096.0 220.024.8 206.096.0 referencemAS -- -- 2.1 2.0 1.3 l.3 0.3 0.2 0.5 0.3 sd -- -- 0.5 0.5 1.3 1.3 0.7 0.5 0.8 0.6 UI -- -- 206.0 204.076.8 76.8 6.0* 4.4 21.6*5.2 dose m.~.S-- -- 2.:1 1.8 2.2 1:.3 1.8 0.8 1.4 0.6 sd -- -- 0.6 0.9 0.5 1..2 1.1 1.3 0.9 0.9 U -- -- 100 100 100 ti0 80 40 80 40 UI -- -- 238.0 178.0218.078.0 145.633.6 115.225.6 dose mAS -- -- 2.1 1.6 1.1 1.1 0.6 0.7 1.2 1.0 sd -- -- 0.7 1.1 1.0 I.0 0.8 0.9 1.1 1.0 U -- -- 100 80 60 fi0 40 40 60 60 UI -- -- 214.0 131.267.2*6:3.624.8*26.4 69.6 62.4 dose mAS -- -- 2.1 1.3 1.2 0.9 0.7 0.9 1.1 0.7 sd -- -- 0.2 0.9 1.2 0.9 1.0 0.9 1.0 0.7 UI -- -- 208.0*107.274.4*5fi.428.8*55.2 66.0 43.2 AS assessment score mAS mean assessment score U percentage of animals with ulcers of all degrees of severity UI ulcer index significance calculation, t-test, p<0.05, versus control Table 3 Area of duodenal ulcer [mmZ]
days post 2 5 8 1.1 21 ulcerogen Group r/n A r/n A r/n A r/n A r/n A
Control 8110 .10.95/5 25.1 4/5 17.5 4/5 13.0 315 14.9 Reference -- -- 3/5 20.9 2/5 21.2 0/5 0 O/S 0 Dose 1 -- -- 4/5 23.4 4/5 12.6 3/5 11.3 2/5 5.9 Dose 2 -- -- 3/5 39.8 2/5 4.7 0/5 0 1/5 3.1 Dose 3 -- -- 4/5 5.7 2/5 8.3 1/5 3.1 0/5 0 Table 4 Area of gastric ulcer [mm2]
days post 2 5 8 14 21 ulcerogen Group r/n A r/n A r/n A r/n A r/n A
Control 5/10 207.33/5 130.93/5 110.00/5 0 1/6 78.5 Reference -- -- 4/5 90.4 2/5 53.4 O/S 0 O/S 0 Dose 1 -- -- 3/5 238.83/5 93.2 1/5 113.11/5 28.3 Dose 2 -- -- 215 64.4 1/5 28.3 0/5 0 115 12.6 Dose 3 -- -- 2/5 28.3 1/5 12.6 0/5 0 O/S 0 r/n number of animals with ulcers of severity degree 2-3/number of animals studied A area (determined for ulcers of severity degree 2-3).
Table 5 _ . Helicobacter pylori (Hp+) [%]
days post 2 5 8 14 21 ulcerogen Group Control 80 100 100 80 80 Reference -- 60 40 20 20 Dose 1 -- 100 60 60 60 Dose 2 -- 60 :l0 40 40 Dose 3 -- 60 20* 0* 0*
* significance calculation, exact Fisher Test, p < 0.05, one-sided, versus control Table 6 Mortality days post 1 2 3 Total uicerogen e/n e/n e/n e/n Group Control 1/30 3/30 x/30 9130 Reference 1/20 0/20 0/20 1/20 Dose 1 0/20 1/20 1/20 2120 Dose 2 1/20 0120 0/20 1120 Dose 3 0/20 0/20 0/20 0/20 e/n number of animals died/total number of animals per group
Claims (6)
Claims
1. Use of an expressed juice, digestive juice or extract of flesh-eating plants for the production of a medicament for the inhibition of protein kinases.
2. Use according to Claim 1, characterised in that the flesh-eating plant belongs to the genus Heliamphora, Saracenia, Darlingtonia, Cephalotus or Nephentes.
3. Use according to Claim 1 or 2, characterised in that the flesh-eating plant belongs to the family Drosera.
4. Use according to at least one of the foregoing Claims, characterised in that the extract is an aqueous extract.
5. Use according to at least one of the foregoing Claims for the control of phototropic bacteria, plasmodia, chlamydiae, trypansomes, staphylococcus bacteria, streptococcus bacteria, candida, mucor and aspergillus.
6. Procedure for treatment of a mammal, including the administration of an expressed juice, digestive juice or extract of flesh-eating plants for the inhibition of protein kinases.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19807285 | 1998-02-23 | ||
| DE19814754A DE19814754A1 (en) | 1998-02-23 | 1998-04-02 | The use of plant extract of meat-eating plants, e.g. Dionacea muscipula |
| DE19814754.6 | 1998-04-02 | ||
| DE19807285.6 | 1998-04-02 | ||
| PCT/EP1999/001176 WO1999042115A1 (en) | 1998-02-23 | 1999-02-23 | Use of a press-juice, digestive juice or extract of carnivorous plants for inhibiting protein kinases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2321783A1 true CA2321783A1 (en) | 1999-08-26 |
Family
ID=26044040
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002321783A Abandoned CA2321783A1 (en) | 1998-02-23 | 1999-02-23 | Use of an expressed juice, digestive juice or extract from flesh-eating plants for the inhibition of protein kinases |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JP2002503699A (en) |
| AU (1) | AU3029799A (en) |
| CA (1) | CA2321783A1 (en) |
| DE (1) | DE19980245D2 (en) |
| WO (1) | WO1999042115A1 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2906818B1 (en) | 2006-10-04 | 2012-04-27 | Plant Advanced Technologies Pat Sas | PROCESS FOR PRODUCING RECOMBINANT PROTEINS USING CARNIVORIC PLANTS |
| SE534483C2 (en) * | 2010-01-26 | 2011-09-06 | Quantum Pharmaceuticals Ltd | New preparation containing extract of Dionaea muscipula for cosmetic treatment of skin |
| DE102011101875A1 (en) * | 2011-05-18 | 2012-11-22 | Merck Patent Gmbh | Extracts from Darlingtonia californica |
| ES2883347T3 (en) * | 2014-06-16 | 2021-12-07 | Codexis Inc | Compositions and procedures to attenuate and prevent intestinal inflammation due to the presence of peptide food antigens in a intestine |
| EP3393502A4 (en) | 2015-12-16 | 2019-06-12 | Nepetx, LLC | Compositions and methods for treating gluten intolerance and disorders arising therefrom |
| CN105995972A (en) * | 2016-05-13 | 2016-10-12 | 晶叶（青岛）生物科技有限公司 | Symbiotic microflora enzyme, application thereof and food prepared from symbiotic microflora enzyme |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE434633C (en) * | 1923-12-28 | 1926-10-01 | Vladislav Mladejovsky Dr | Process for the preparation of a Drosera extract which contains the active ferments and is particularly active in arteriosclerosis |
| DE3573069D1 (en) * | 1985-07-31 | 1989-10-26 | Keller Helmut | Medicine for the treatment of chronical inflammatory enterocolitis |
| ATE46443T1 (en) * | 1985-07-31 | 1989-10-15 | Helmut Dr Med Keller | MEDICATIONS FOR THE TREATMENT OF HERPES. |
| DE3619281A1 (en) * | 1986-06-07 | 1987-12-10 | Helmut Dr Med Keller | MEDICINES FOR TREATING MALIGNERS AND CHRONIC DISEASES |
| DE3844244A1 (en) * | 1988-12-19 | 1990-06-21 | Florian Draenert | Compounds having antibiotic, cytostatic and/or antiviral action derived from carnivorous plants |
-
1999
- 1999-02-23 CA CA002321783A patent/CA2321783A1/en not_active Abandoned
- 1999-02-23 AU AU30297/99A patent/AU3029799A/en not_active Abandoned
- 1999-02-23 WO PCT/EP1999/001176 patent/WO1999042115A1/en active Application Filing
- 1999-02-23 DE DE19980245T patent/DE19980245D2/en not_active Expired - Fee Related
- 1999-02-23 JP JP2000532129A patent/JP2002503699A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| WO1999042115A1 (en) | 1999-08-26 |
| JP2002503699A (en) | 2002-02-05 |
| AU3029799A (en) | 1999-09-06 |
| DE19980245D2 (en) | 2001-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20130142696A (en) | Composition comprising protaetia brevitarsis for preventing and treating inflammatory disorder | |
| US20110059193A1 (en) | Method for obtaining an extract of cranberry marc that can be used in particular in the prevention and treatment of conditions such as caries, gingivitis or sore throats | |
| JP2009029772A (en) | Enzyme-treated royal jelly, and antioxidant, moisturizer, skin fibroblast proliferation promoter, hypotensive, fatigue-relieving agent and calcium absorption promoter, each containing the same | |
| KR100815171B1 (en) | Cyclooxygenase Inhibitor, and Food and Beverage containg the Same | |
| KR100964906B1 (en) | Pharmaceutical composition for treating or preventing an inflammatory disease comprising licarin E | |
| CA2321783A1 (en) | Use of an expressed juice, digestive juice or extract from flesh-eating plants for the inhibition of protein kinases | |
| KR101219551B1 (en) | Composition of comprising bee-venoms for Neuroprotective properties | |
| KR100700480B1 (en) | Composition comprising the extract of Phellodendron amurensis for preventing and treating fracture | |
| JPH07258103A (en) | Antithrombosis agent and its preparation | |
| JPH0427964B2 (en) | ||
| RU2593339C2 (en) | Natural drug composition | |
| JP3547835B2 (en) | Prophylactic / therapeutic agent for throat inflammation and hemolytic toxin and oral composition containing the same | |
| KR20200046772A (en) | Composition comprising Locusta migratoria extract for preventing or treating Neuritis | |
| US20170128513A1 (en) | Cinnamomum burmanii extract, extraction process and its use as proton pump down-regulator, enzyme inhibitor, and mucoprotector | |
| KR102123030B1 (en) | A composition for improving diabetes or for use of antioxidant comprising yeast extract and a method for preparing the yeast extract | |
| JPS5821638B2 (en) | glycyrrhizinsanno | |
| JP3916686B2 (en) | Glycerophosphate dehydrogenase inhibitor | |
| JP2772552B2 (en) | Preventive agent for dental caries and periodontal disease | |
| JP3183754B2 (en) | Arteriosclerosis inhibitor and composition containing the same | |
| KR100514315B1 (en) | Composition containing cordyceps sp. or paecilomyces japonica extract for inhibiting activity of matrix metalloproteinase | |
| JPH09286737A (en) | Glycerophosphate dehydrogenase inhibitor | |
| TW202200179A (en) | Composition for inhibiting phosphodiesterase 5 activity or improving erectile function by using a plant of the genus Phyllanthus | |
| KR20210011051A (en) | Composition comprising Locusta migratoria extract for preventing or treating Neuritis | |
| NO170681B (en) | ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY ACTIVE GUANIDINASPARAGINALS | |
| KR20050100719A (en) | Composition comprising the extract of chaga mushroom having anti-inflammatory activity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |