WO2002047697A2 - Fractions of snake blood, their preparation and use as therapeutic agents - Google Patents
Fractions of snake blood, their preparation and use as therapeutic agents Download PDFInfo
- Publication number
- WO2002047697A2 WO2002047697A2 PCT/EP2001/014727 EP0114727W WO0247697A2 WO 2002047697 A2 WO2002047697 A2 WO 2002047697A2 EP 0114727 W EP0114727 W EP 0114727W WO 0247697 A2 WO0247697 A2 WO 0247697A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fractions
- blood
- serum
- snake
- preparation
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/58—Reptiles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- the present invention refers to fractions of snake blood and to their preparation and use as therapeutic agents.
- the active principles present in snake blood show therapeutic activity on cell lines
- Chemotherapeutics are usually classified according to their chemical structure or their biological activity (see “The Pharmacological Basis of Therapeutics” by A. Goodman and M. Gilman, Pergamon Press, New York, 1990). In most cases anti tumour drugs act on the cell life cycle blocking the cell growth.
- the chemotherapeutics classification includes also natural compounds or derivatives thereof (as taxol or vincristine) which are capable to bind to tubulin and therefore inhibits cell mitosis.
- chemotherapeutic treatment of tumours it is self evident the importance of making available new drugs having anti tumour activity.
- the invention refers to fractions of snake blood obtained by centrifugation of snake blood and their therapeutic use.
- Fig. 1 is a diagram showing the optical density of various cell lines and controls after incubation with P Serum 1:100 for 1 h.
- Fig.: 2A reports the optical density of A431 cells cultures treated with various dilution of P Serum incubated at 56°C for 10' or 1 h at 37°C.
- Fig. 2B reports the optical density of A431 cells cultures treated with various dilution of P Serum incubated at 56°C for 10' or 3 h at 37°C.
- Fig. 3 shows the optical density of various cell lines grown in the presence of P
- Fig. 4A shows the number of viable cells per well of A431 cells treated with P
- Fig. 4B shows the number of viable cells per well of U373-MG cells treated with P
- Fig.: 4C shows the number of viable cells per well of HF cells treated with P
- Fig. 4D shows the number of viable cells per well of CVEC cells treated with P
- Fig. 5 is the picture of the electrophoresis on SDS-PAGE 12% of the supernatant obtained in Example 1.
- the present invention makes available new compounds having anti tumour activity consisting of fractions of snake blood.
- fractions means the supernatant obtained by centrifugation of the snake blood (hereinafter P Serum) or the precipitate
- pellet obtained by centrifuging solution obtained treating the above said P Serum with a ethanol/water solution (such treatment being possibly repeated one or more times).
- the snake blood used as starting product for the centrifugation is the blood of snakes of genus pythons.
- aliquots of snake blood are collected by intracardiopuncture and are centrifuged, the supernatant (P Serum) is collected and frozen.
- the P Serum obtained as above described is treated with an ethanol/water solution and the so obtained solution is centrifuged, the supernatant is now eliminated collecting the precipitate (the operation can be repeated more then once) the precipitate remaining after the last centrifugation is collected and tested.
- the centrifugation of the snake blood is preferably performed at 3000 rpm for 5 minutes while the centrifugation of the ethanol/water solution is preferably performed at 15000 rpm for 30 minutes.
- the composition of the ethanol/water solution is preferably comprised between
- the solution is centrifuged at 15000 rpm for 30 minutes; the supernatant is eliminated and the pellet is collected.
- the pellet was resuspended in a solution of ethanol and water (50%/50%) and the procedure was repeated three folds.
- Serum P The specific anti tumour effect of Serum P (as obtained in example 1) was determined performing tests on various cell lines, in particular:
- MCF7 human breast carcinoma
- B16 murine melanoma
- the cell viability was evaluated with the sulphurodamine B test according to
- Serum P at different concentration (1:25, 1:50; 1:100, 1:200) is poured in the wells in the absence or in the presence of FCS, the incubation time of the tested samples was 1h and 18 h respectively.
- microtiter plate was kept at 4°C for 1.5 h and then washed with water (6 folds).
- P Serum is very toxic on all the tested cell lines (tumour and non tumour) at dilution 1:25 and 1:50. This toxicity is evident already after 1 h incubation (see
- Table 1 Effect of P Serum on the viability of tumor and non-tumor cell lines (1 h stimulation)
- P Serum up to 1 :100 show a different effect on tumour cells in respect of non tumour cells.
- the P Serum shows low or absent toxicity on endothelial cells, a relatively high toxicity on B16 cells and a very high toxicity on other tumour cells (A431, MCF-7 and U373) (see Fig. 1).
- the effects of P Serum on cell viability are absent when the P Serum is incubated at 56°C for 10 minutes (see Fig. 2 A and B): in the figures are reported the effects on cell line A431 , and similar effects are observed in respect of other cell lines tested (data not shown).
- the anti tumour activity of the pellets was tested on A 431 (human squamous carcinoma cell line) and on the cell line of bovine endothelial microcirculus (CVEC) as non tumour control.
- the cell viability was measured as said above using the Sulphurodamine B test.
- the anti tumour activity of the pellets obtained according to example 3 was tested on A431 and CVEC as control.
- the cell viability was tested using the sulphurodamine B test as above said.
- the desired product is a protein of about 33 kDa having S-S bridges probably in the form of a dimer or oligomer.
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2002217110A AU2002217110A1 (en) | 2000-12-15 | 2001-12-14 | Fractions of snake blood, their preparation and use as therapeutic agents |
EP01270337A EP1349557A2 (en) | 2000-12-15 | 2001-12-14 | Fractions of snake blood, their preparation and use as therapeutic agents |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITFI2000A000251 | 2000-12-15 | ||
IT2000FI000251A IT1319158B1 (en) | 2000-12-15 | 2000-12-15 | PHARMACEUTICAL COMPOSITION WITH ANTI-TUMOR ACTIVITY CONSTITUTING SNAKE BLOOD COMPONENTS AND METHOD FOR THEM |
Publications (3)
Publication Number | Publication Date |
---|---|
WO2002047697A2 true WO2002047697A2 (en) | 2002-06-20 |
WO2002047697A3 WO2002047697A3 (en) | 2002-11-07 |
WO2002047697A8 WO2002047697A8 (en) | 2003-03-20 |
Family
ID=11442012
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2001/014727 WO2002047697A2 (en) | 2000-12-15 | 2001-12-14 | Fractions of snake blood, their preparation and use as therapeutic agents |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP1349557A2 (en) |
AU (1) | AU2002217110A1 (en) |
IT (1) | IT1319158B1 (en) |
WO (1) | WO2002047697A2 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007085879A1 (en) * | 2006-01-26 | 2007-08-02 | Marina Ziche | Compositions containing substances obtained from the blood and other biological fluid of snake : methods for their preparation and their use |
ITFI20110168A1 (en) * | 2011-08-05 | 2013-02-06 | Filippis Vincenzo De | PROTEINS WITH APOPTOTIC AND ANTITELOMERASIC ACTIVITY ON CANCER CELLS, THEIR PREPARATION AND USE. |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4012502A (en) * | 1973-09-21 | 1977-03-15 | Buren Philpot V Jun | Snake venom inhibitor material and method of purification |
WO1999029726A1 (en) * | 1997-12-05 | 1999-06-17 | Hsc (Pla) Pty. Ltd. | Phospholipase inhibitor |
-
2000
- 2000-12-15 IT IT2000FI000251A patent/IT1319158B1/en active
-
2001
- 2001-12-14 WO PCT/EP2001/014727 patent/WO2002047697A2/en not_active Application Discontinuation
- 2001-12-14 EP EP01270337A patent/EP1349557A2/en not_active Withdrawn
- 2001-12-14 AU AU2002217110A patent/AU2002217110A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4012502A (en) * | 1973-09-21 | 1977-03-15 | Buren Philpot V Jun | Snake venom inhibitor material and method of purification |
WO1999029726A1 (en) * | 1997-12-05 | 1999-06-17 | Hsc (Pla) Pty. Ltd. | Phospholipase inhibitor |
Non-Patent Citations (2)
Title |
---|
THWIN M ET AL: "SNAKE ENVENOMATION AND PROTECTIVE NATURAL ENDOGENOUS PROTEINS: A MINI REVIEW OF THE RECENT DEVELOPMENTS (1991-1997)" TOXICON, ELMSFORD, NY, US, vol. 36, no. 11, 1998, pages 1471-1482, XP001024676 ISSN: 0041-0101 * |
THWIN MM ET AL: "Recombinant antitoxic and antiinflammatory factor from the nonvenomous snake Python reticulatus: phospholipase A2 inhibition and venom neutralizing potential" BIOCHEMISTRY, AMERICAN CHEMICAL SOCIETY. EASTON, PA, US, vol. 39, no. 31, 8 August 2000 (2000-08-08), pages 9604-9611, XP002179017 ISSN: 0006-2960 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007085879A1 (en) * | 2006-01-26 | 2007-08-02 | Marina Ziche | Compositions containing substances obtained from the blood and other biological fluid of snake : methods for their preparation and their use |
ITFI20110168A1 (en) * | 2011-08-05 | 2013-02-06 | Filippis Vincenzo De | PROTEINS WITH APOPTOTIC AND ANTITELOMERASIC ACTIVITY ON CANCER CELLS, THEIR PREPARATION AND USE. |
WO2013021339A1 (en) * | 2011-08-05 | 2013-02-14 | Marina Ziche | Proteins with pro-apoptotic and antitelomerase activity on cancer cells, their preparation and use |
Also Published As
Publication number | Publication date |
---|---|
WO2002047697A3 (en) | 2002-11-07 |
ITFI20000251A1 (en) | 2002-06-15 |
EP1349557A2 (en) | 2003-10-08 |
WO2002047697A8 (en) | 2003-03-20 |
IT1319158B1 (en) | 2003-09-26 |
AU2002217110A1 (en) | 2002-06-24 |
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