WO1995014489A1 - Composition comprenant des composes utilises comme agent therapeutique - Google Patents

Composition comprenant des composes utilises comme agent therapeutique Download PDF

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Publication number
WO1995014489A1
WO1995014489A1 PCT/SE1994/001112 SE9401112W WO9514489A1 WO 1995014489 A1 WO1995014489 A1 WO 1995014489A1 SE 9401112 W SE9401112 W SE 9401112W WO 9514489 A1 WO9514489 A1 WO 9514489A1
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Prior art keywords
phim
group
cells
rats
ions
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PCT/SE1994/001112
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English (en)
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Johan De Faire
Ragnvald Lindblom
Karl-Erik Arfors
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Phairson Medical Inc.
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Priority to AU10821/95A priority Critical patent/AU1082195A/en
Publication of WO1995014489A1 publication Critical patent/WO1995014489A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6402—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • Composition comprising compounds for use as therapeutical agent .
  • the present invention relates to new pharmaceutical compounds or compositions of non-immunogenic proteinaceous substances which have recognition, targeting and enzymatic activity and which in tests surprisingly have proven to provide a spectrum of different properties depending on salt compiexing compounds and therefore will be useful for cure of severe diseases.
  • these substances function in an immune defense like manner, i.e. they seem to be able to distinguish between various particles and soluble matter regarded to be " normal” or "abnormal", respectively, in a particular environment or. differently expressed, seem to be able to recognizing, targeting and destroying divergent cells. It should, however, be emphasized that the invention is not intented to restricted by any hypothetic mode of action expressed in the present specification.
  • WO 85/04809 disclosed the uses of enzymes from Antarctic krill as a digestion promotor.
  • EP-A1- 0170115 discloses the use of krill as a thrombus dissolvent.
  • Enzymes and enzyme mixture derived from Antarctic krill have been isolated and extensively studied as regards their biological and biochemical propertiesNarious procedures for isolating these enzymes have been developed. See e.g. Anheller J.E.. Hellgren L.. Karlstam B.. and Vinsent J. ( 1989): "Biochemical and biological profile of a new enzyme preparation from Antarctic krillC E. superba): Axeisen N.H.. Kroll J. and Weeke B. (1973): " A manual of quantitative immunoelectrophoresis: methods and applications:. Scan. J. Immunol. 2. Suppl.: Bucht A. and Karlstam B.
  • Our immune system is always on guard and active and its main functions are to protect and preserve the integrity of our body by taking care of foreign invaders, old and wom-out cells, sick cells and soluble matters that are regarded to be abnormal in the specific environment.
  • Phagocytes, granulocytes. macrophages. NK-cells. CRP. complement system. lymphocytes. T-cells. B-cells etc. are our " immune defense system" and their mode of action for protection and preservation is to. recognize, targeting and destroying of foreign invaders, old and worn-out cells, sick cells or abnormal cells.
  • the whole immune defense system works and interact in a complex interplay for optimal protection of our body.
  • the sign of an activated immune defense system can be seen as an inflammation which is caused be leaking "toxins" and overproduction of TNF. IL-2 etc.
  • the inflammation is no harmful as long as it is kept at a low level.
  • the immune defense system gets over-activated in such a way that the patient is dying of to large produced
  • TNF.IL-2 etc. or to large amount of necrotic tissue, i.e. in the case of cancer.
  • SUBSTITUTE SHEET The microbes may disguise themselves to irrecognition for the immuno-cells or they may enter into cells, even immuno-cells. and live, multiply, and change the memory of their host-cells. Some cancers and AIDS and the severe complication of opportunistic infections of these diseases are examples of such microbial ( mis-) behavior.
  • the pathogenesis of a disease is thus dependent on how well the immune defense system is functioning and normally it is in perfect balance for its tasks. However the pathogenesis of a diseases is also dependent of the influence of the used drugs on the immune defense system.
  • drugs are the main cause of temporary and permanent suppression of the immune defense system and much attention in pharmaceutical research is put in this direction to create drugs which attack the "sick cells" and not the healthy cells.
  • the optimal drug would be a drug that acts in harmony and symbiotic interplay with the immune defense system and with recognition, targeting to the place for the sickness and destroying the sick cells without adverse reactions.
  • the present invention provides novel pharmaceutical compounds which seem to have said properties.
  • the means by which this is achieved involves the use of one or more proteolytic enzymes with endo and exo peptidase properties which have been isolated from Antarctic krill(Euphasia superba) combined with suitable "salt complexing compounds". As mentioned above, such kind of enzymes are known as such, and have also been suggested for a few pharmaceutical uses.
  • the water phase containing less than 2% fat. was diluted with distilled water to a conductivity of 1.5 S/cm. and then applied to a Q-Sepharose column( Pharmacia. Sweden).equilibrated with 0.05M TRIS-buffer at pH:5.5
  • IM sodium chloride buffer was added to the Q-Sepharose column at a gradient over 20 column volumes. Most of the proteins in the water phase were eluted in the void peak and the proteins were collected from approximately 50% to 95% sodium chloride content.
  • the collected protein factions were desalted on a G-25 Sephadex column(Pharmacia Sweden) and then reapplied to an equilibrated Q-Sepharose column and rechomatographed as above.
  • Trypsin 1 All the proteins hydrolyse Tyrosine-Arginine-Methyl-Ester(TAME)(Sigma Chemical Corp.. USA) and will therefore be designated Trypsin 1. Trypsin 2 resp. Trypsin 3. Henceforth in the text. Trypsin 1 will be identical to PHIM 101 and the mixture of Trypsin 1-2-3 will be indentical to PHIM 103.
  • the molecular weight for Trypsin 1 is approx. 29kDaltons and 31kDaltons for both Trypsin 2 and 3. determined by SDS-Page.
  • Trypsin 1.2 and 3 have isoelectric focusing between pi 3.5-4.5.
  • pH optima are at pH 8-8.5 for all the Trypsins.
  • Trypsin 1 express both exo- and endo-peptidase activity in the same molecule and it may liberate both free amino acids and peptide fragments from denatured milk casein.
  • Trypsin 2 and 3 are traditional endo-peptidases and may not liberate free amino acids from casein when added equivalent TAME Units as for trypsin 1.
  • PHIM 101 3 mg is dissolved m 3 mi of 0.2 M NaH2P04 at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule.
  • SUBSTITUTE SHEET The characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 mm. and 180 min. and expressed as TAME units(TU).
  • the second method is to add 0.15 M NaCL and measure at 0 mm.. 60 min. and 180 min.
  • the third method is to add ATP instead of NaCl and measure after 0 min.. 60 min. and 180 mm.
  • exo-peptidase activite was also determined with following method:
  • PHIM 101.1 mg ml was incubated for 6 hours at 30°C with 1% casein preparation in order to liberate free amino acids.
  • PHIM 101 3 mg is dissolved in 3 ml of 0.2 M TRIS-HC1 at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule. The characterization of the enzymatic activity is performed by reconstimted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU). The second method is to add 0.15 M NaCL and measure at 0 min.. 60 min. and 180 min. The third method is to add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • PHIM 101.1 mg/ml was incubated for 6 hours at 30°C with 1% casein preparation in order to liberate free amino acids.
  • PHIM 101 3 mg is dissolved i-n 3 ml of 0.2 M NaCl at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule. The characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU). The third method is to add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • exo-peptidase activite was also determined with following method:
  • PHIM 101.1 mg/ml was incubated for 6 hours at 30°C with 1% casein preparation in order to liberate free amino acids.
  • PHIM 101 3 mg is dissolved in 3 ml of 0.2 M Glucose amine at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule. The characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME umts(TU). The second method is to add 0.15 M NaCL and measure at 0 min.. 60 min. and 180 min. The third method is co add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • exo-peptidase activite was also determined with following method:
  • PHIM 101.1 mg/ml was incubated for 6 hours at 30°C with 1% casein preparation in order to liberate free amino acids.
  • PHIM 101 3 mg is dissolved in 3 ml of 0.2 M ATP at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule.
  • the characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU).
  • the second method is to add
  • PHIM 103 3 mg is dissolved in 3 mi of 0.2 M NaH2P04 at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule. The characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU). The second method is to add 0.15 M NaCL and measure at 0 min.. 60 min. and 180 min. The third method is to add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • PHIM 103 3 mg is dissolved in 3 ml of 0.2 M TRIS-HCl at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule.
  • the characterization of the enzymatic activitv- is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU).
  • the second method is to add 0.15 M NaCL and measure at 0 min.. 60 min. and 180 min.
  • the third method is to add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • PHIM 103 3 mg is dissolved in 3 ml of 0.2 M NaCl at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule. The characterization of the enzymatic activity is performed by reconstimted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 mm. and 180 mm. and expressed as TAME un ⁇ ts(TU). The third method is add ATP instead of NaCL and measure after 0 mm.. 60 min. and 180 min.
  • PHIM 103 3 mg is dissolved in 3 ml of 0.2 M Glucose amine at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule.
  • the characterization of the enzymatic activity is performed by reconstituted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME u ⁇ its(TU).
  • the second method is to add 0.15 M NaCL and measure at 0 min.. 60 min. and 180 min.
  • the third method is to add ATP instead of NaCL and measure after 0 min.. 60 min. and 180 min.
  • PHIM 103 3 mg is dissolved in 3 ml of 0.2 M ATP at pH 7.4. The solution is equilibrated during 30 minutes. The solution was lyophilized in ampoule.
  • the characterization of the enzymatic activity is performed by reconstimted the product in sterile water and determined by TAME method at 247 nm at 0 min. 60 min. and 180 min. and expressed as TAME units(TU).
  • the second method is to add
  • Figure 1 shows the weight of the rats at the treatment day and 5 days later.
  • Figure 2 shows the tumor growth from the start of the treatment and 5 days later.
  • Figure 3 shows the tumor reduction compared with the control group.
  • Figure 4 shows the surface of the necrotic part of the tumors in the treatment groups.
  • Figure 5 shows the weight of the tumors in each group compared with the control.
  • Figure 6 shows the tumor size in each group at the day for death.
  • Figure 7 shows the median survival of the control group vs treatment groups.
  • Figure 8 shows the tumor necrotic surface compared with the whole tumor.
  • the purpose of the study was to investigate the efficacy and usefulness of PHIM 101 combined with Phosphate buffer.
  • Tris-HCl buffer Calcium chloride and sterilized water on treatment of Yoshida Sarcoma in rats.
  • the rats were randomized in 5 groups with 5 animals in each group.
  • control group without treatment was coded as group zero.
  • Group 1 was treated with PHIM 101 in Phosphate buffer.
  • Group 2 was treated with PHIM 101 and TRIS buffer.
  • Group 4 was treated with PHIM 101 in sterile water.
  • test drug PHIM 101 was injected subcutaneous to white Wistar rats with implanted Yoshida Sarcoma.
  • PHIM 101 was used as repeated doses of 5 mg/kg B/W. The first dose was injected the fifth day after implantation and the second doses was injected 24 hours later. The rats were sacrificed 4 days after the second and last injection.
  • the size of the mmors were measured before and 4 days after the last treatment and compared with the control as well as the weight of the tumor.
  • the tumor growth during the last five days was enormous with around 1700% in the control group.
  • TNF Tumor Necrosis Factor
  • PHIM 101 is the code name for the protein with both endo and exo peptidase activity in the same molecule origin from Krill.
  • the protein recognize and target sick cells/cancer cells and by the enzymatic properties destroying the targeted cells.
  • the second working hypothesis is that the protein is capable for recognize the same signals as our own immune defense system do. to attack all foreign invaders as well as sick cells.
  • CRP C-Reactive protein
  • Cancer cells as well as microbial infected cells are sick or divergent in the sense that they express fragments of tumor proteins on the cell surface.
  • the treatment started when the implanted mmor cells had grown up to a size of 10x10 mm.
  • the rats were sacrificed 4 days after the treatment was finished and the size of the tumor were measured, the size of the necrotic surface were measured and the weight of the mmors noted.
  • the treatment of the rats started when the size of the mmors were 10x10mm..
  • the animal were divided in 5 groups.
  • Group 0. was the control group of 5 rats without treatment.
  • Group 1 Consist of 5 rats and were treated with 5mg PHIM 101 in Phosphate buffer/ kg BAV subcutaneous about 4 cm from the mmor. The rats received 2 injection with 24 hours differences.
  • the mmors are growing very fast and the increase of the size of the tumors from the treatment day to the rats are sacrificed 5 days later are for the control group about
  • necrotic surface was biggest in group 1 compared with the rest of the treatment groups and the differences was so much as 58% bigger for group 1 compared with group 4 and 46% bigger then group 2 and 3 and 46% ( Fig. 4).
  • the weight of the treated tumors are related to the tumor response and logical the best group nr. 2 with an efficacy of 58% had also the smallest weight of the mmors and also from the logical point of view the most heavy tumors are from the control group and the weight of the mmors in group 1 are closer to the control then the rest of the treatment groups(Fig. 5).
  • rats in group 1 with large tumor necrotic surface looks very sick at day 4 and showed typical TNF and necrotic tissue toxicity.
  • the number of rats in each group was the same and enough to draw conclusion from.
  • TRIS buffer showed the best mmor results is probably that the TRIS molecule us such stabilize, interact and establish a complex with the PHIM molecule as PHIM 101 is very negative charged and TRIS is weak basic. Furthermore the salt complex of PHIM 101 and TRIS could be regarded as the state of equilibrium where both exo- and endo-peptidase actvities are expressed in
  • TRIS in this case is more than a buffer.
  • TRIS a chemicai compound also change the properties of PHIM 101 in such a way that we will get a kind of "slow release system" and that is beneficial for the efficacy and act as a slow release of PHIM 101.
  • PHIM 101 Tris probably are homing to the cells at the surface of the mmors and stop the angiogenesis which prevent spreading of new cells as well as metastasis.
  • PHIM 101 in Phosphate buffer is very activated and are probably homing to the central part of the tumor direct and therefore is no more PHIM 101 available for slow down the growth of the mmor.
  • the degradation of the tumor cells in the central part of the tumor is so strong that the large amount of necrotic tissue cannot be adsorbed by the body and therefore an over production of TNF as well as toxic products from the necrotic cells produce severe toxic symptoms on the animals.
  • PHIM 101 with Calcium Chloride and sterile water are regarding to the mmor efficacy between the group with PHIM 101 and Phosphate buffer and TRIS buffer.
  • TRIS have an influence on both the recognition, targeting and destroying properties of PHIM is must be very important to select right "buffer system" for different purpose. What is optimal for treatment of cancer may by different for microbial infected cells.
  • buffer system together with i.e. proteins and enzymes etc. is to keep a stable pH of the solution and to keep a good stability of the solution. Buffer system do not interfere with the active compound as such.
  • buffer system together with PHIM 101 cannot be regarded as true buffer systems they must be regarded as an activator or deactivator of the PHIM
  • the rats were randomized in 4 groups.
  • PHIM 101 is the code name for the protein with both endo and exo peptidase activity in the same molecule origin from Krill.
  • the protein recognize and target sick cells/cancer cells and by the enzymatic properties destroying the targeted cells.
  • the second working hypothesis is that the protein is capable for recognize the same signals as our own immune defense system do. to attack all foreign invaders as well as sick ceils.
  • CRP C-Reactive protein
  • Cancer cells as well as microbial infected cells are sick or divergent in the sense that they express fragments of mmor proteins on the cell surface.
  • the white powder was reconstituted in:
  • the animal were divided in 5 groups.
  • Group 0 was the control group of 5 rats without treatment.
  • Rat nr. 1-5 ascites and metastasis in all the rats.
  • Rat nr. 2-4. dilute ascites. but not as much as the control.
  • Phosphate buffer group but less ascites mmors.
  • the rats were randomized in 2 groups with 6 rats in control group and 5 rats in the treatment group.
  • control group without treatment was coded as group zero.
  • Group 1 was treated with PHIM 101 in Phosphate buffer.
  • test drug PHIM 101 was injected subcutaneous to white Wistar rats with implanted Yoshida Sarcoma.
  • PHIM 101 was used as repeated doses of 5 mg/kg BAV once a day.
  • the first dose was injected the fifth day after implantation. The study was terminated when all the rats were dead
  • the size of the mmors were measured at the first treatment day and when each rat is dead.
  • the necrotic surface was measured.
  • the median survival was 5 days in the treatment group and 8 days in the control group.
  • the mmor growth during the five days was 748% in the treatment group and during the 8 days in the control group the mmor growth was 2095%.
  • the degree of necrotic surface was 11.6% in the treatment group and 7.7% in the control group.
  • PHIM 101 is the code name for the protein with both endo and exo peptidase activity in the same molecule origin from Krill.
  • the protein recognize and target sick cells/ cancer cells and by the enzymatic properties destroying the targeted cells.
  • the second working hypothesis is that the protein is capable for recognize the same signals as our own immune defense system do. to attack all foreign invaders as well as sick cells.
  • CRP C-Reactive protein
  • Cancer cells as well as microbial infected cells are sick or divergent in the sense that they express fragments of mmor proteins on the cell surface.
  • the design of the study is a randomized comparative study with injection of PHIM
  • the treatment started when the implanted tumor cells had grown up to a size of 10x10 mm.
  • the size of the tumor were measured and the size of the necrotic surface were measured.
  • the white powder was reconstituted in:
  • the treatment of the rats started when the size of the mmors were 10x10mm..
  • the animal were divided in 2 groups.
  • Group 0. was the control group of 5 rats without treatment.
  • Group 1 Consist of 6 rats and were treated with 5mg PHIM 101 in Phosphate buffer/ kg B/W subcutaneous about 4 cm from the mmor once a day.
  • the tumors are growing very fast and the increase of the size of the tumors from the treatment day to the rats are dead median value of 5 days for the rats in the treatment groups was 765% and for the untreated rats 2095%( Fig. 6) he median survival of the rats in the treatment group was 5 days and for the rats in the control group 8 days despite much bigger tumors(Fig. 7).
  • the surface of the necrotic field is bigger for the treated rats as expected.
  • the reason for the shorter survival depends on to good efficacy and necrosis of a fast growing large mmors which leads to ove ⁇ roduction of TNF and leakage of toxic tumor necrotic tissue.
  • PHIM 101 and TRIS-HCl in concentration of 1 ug/ml-1000ug'ml was added to a standard in vitro test with living cells and Hantaan virus, the degree of replication was measured of the virus.
  • the inhibition of virus was between 25- 75%.
  • TBC 5-Mvcobacterium tuberculosis
  • PHIM 101 and TRIS-HCL in concentration of 1-500 ug/ml was added to fluorescein labelled TBC bacte ⁇ a and macrophages and the amount of surviving cells was measured.
  • test compound destroyed only the TBC containing macrophages and the amount of TBC cells decreased between 10-85% depending on the concentration of PHIM
  • Red cells were produced in culture with 10-20% malaria (Plasmodium falciparum) containing cells.
  • the red membrane were stained with Carboxyfluorescein 2mg/ml ( Molecular Probes. Eugene. OR USA) and the maiarial parasites within the red cells with Ethidium bromide and in some experiments with Thiazole orange. ( Becton
  • PHIM 101 Depending on the specific salt-complex formed between PHIM 101 and added molecules/ ions, the activity balance of expressed exo- and endo-peptidase activity may be shifted from one activity to an other. Together with phospate and glucose amine solutions. PHIM 101 preferently expresses endo-peptidase activity. In water solutions. PHIM 101 expresses mainly exo-peptidase activity. PHIM 101 in TRIS-HCl solutions could be regarded as the state of equilibrium where both exo- and endo ⁇ peptidase activities are expressed in comparable ratio which is very important for breaking down sick cells and tissues in an optimal way.

Abstract

L'invention porte sur une composition comprenant des enzymes non immunogènes du type exo et endopeptidases isolées à partir du krill Euphasia superba de l'antarctique et des composés de complexant des sels utilisables comme agents thérapeutiques.
PCT/SE1994/001112 1993-11-22 1994-11-22 Composition comprenant des composes utilises comme agent therapeutique WO1995014489A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU10821/95A AU1082195A (en) 1993-11-22 1994-11-22 Composition comprising compounds for use as therapeutical agent

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Application Number Priority Date Filing Date Title
SE9303899A SE9303899D0 (sv) 1993-11-22 1993-11-22 Läkemedel
SE9303899-0 1993-11-22

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006022947A1 (fr) * 2004-07-22 2006-03-02 University Of Chile Proteine et sequence d'acides nucleiques codant une enzyme a activite de type trypsine adaptee au froid derivee du krill
WO2008054293A1 (fr) * 2006-11-03 2008-05-08 Salutary Care Limited Composition, utilisation de ladite composition pour le traitement de maladies et d'états systémiques, et produit contenant ladite composition
CN109030680A (zh) * 2018-09-13 2018-12-18 山东师范大学 一种南极磷虾中泼尼松龙、醛固酮、睾酮和雌二醇的提取以及检测方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984001715A1 (fr) * 1982-10-25 1984-05-10 Hellgren Lars G I Composition enzymatique pour nettoyage therapeutique et/ou non therapeutique, utilisation et preparation de la composition
WO1989010960A1 (fr) * 1988-05-05 1989-11-16 Pharmacia Ab Procede de modification de proteines, peptides et/ou lipides au moyen d'enzymes provenant d'euphauciaces
WO1993024142A1 (fr) * 1992-05-22 1993-12-09 Phairson Medical Ab Nouvelles utilisations pharmaceutiques des enzymes du krill

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984001715A1 (fr) * 1982-10-25 1984-05-10 Hellgren Lars G I Composition enzymatique pour nettoyage therapeutique et/ou non therapeutique, utilisation et preparation de la composition
WO1989010960A1 (fr) * 1988-05-05 1989-11-16 Pharmacia Ab Procede de modification de proteines, peptides et/ou lipides au moyen d'enzymes provenant d'euphauciaces
WO1993024142A1 (fr) * 1992-05-22 1993-12-09 Phairson Medical Ab Nouvelles utilisations pharmaceutiques des enzymes du krill

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DIALOG INFORMATION SERVICES, File 347, Japio, Dialog Accession No. 01854319, KAO CORP, "Anti-inflammatory Agent"; & JP,A,61 068 419, (08-04-1986). *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006022947A1 (fr) * 2004-07-22 2006-03-02 University Of Chile Proteine et sequence d'acides nucleiques codant une enzyme a activite de type trypsine adaptee au froid derivee du krill
WO2008054293A1 (fr) * 2006-11-03 2008-05-08 Salutary Care Limited Composition, utilisation de ladite composition pour le traitement de maladies et d'états systémiques, et produit contenant ladite composition
CN109030680A (zh) * 2018-09-13 2018-12-18 山东师范大学 一种南极磷虾中泼尼松龙、醛固酮、睾酮和雌二醇的提取以及检测方法

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AU1082195A (en) 1995-06-13

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