Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/08—Tripeptides
  • C07K5/0815—Tripeptides with the first amino acid being basic
• • 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
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides
  • C07K5/06086—Dipeptides with the first amino acid being basic
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/50—Cyclic peptides containing at least one abnormal peptide link
  • C07K7/54—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring
  • C07K7/60—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring the cyclisation occurring through the 4-amino group of 2,4-diamino-butanoic acid
  • C07K7/62—Polymyxins; Related peptides
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/64—Cyclic peptides containing only normal peptide links
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
  • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
  • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P20/00—Technologies relating to chemical industry
  • Y02P20/50—Improvements relating to the production of bulk chemicals
  • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

• This invention relates to methods for preparing novel protected peptide intermediates from polymyxin B or from related lipopeptide antibiotics, which are used to readily prepare new families of antibiotics that have potent activity against gram-negative and gram-positive bacteria.
• Polymyxin B and the related colistin have been used in humans but their use has been previously restricted because of toxicity and the availability of the other less toxic and previously effective antibiotics cited above (Ref. 1). Polymyxin B and colistin are usually administered by intravenous or intramuscular dosing.
• Colistimethate sodium is a water soluble salt of colistin/formaldehyde/bisulfite and has been of particular therapeutic value in acute and chronic urinary tract infections caused by strains of Pseudomonas aeruginosa (PDR Generics).
• Polymyxin B sulfate is the drug of choice in the treatment of infections of the urinary tract, meninges, and bloodstream caused by susceptible strains of P. aeruginosa (PDR Generics). Aerosolized polymyxin B is an important component of therapeutic regimens used in the management of pseudomonal lung infections, characteristically found in cystic fibrosis (Ref. 2). Some renal toxicity was observed with recommended dosing of polymyxin B in patients.
• the total synthesis of polymyxin B and four analogs was accomplished by a combination of solid phase peptide syntheses to obtain linear structures followed by removal from the resin and condensation in solution at high dilution to obtain the cyclic peptides (Ref. 7).
• the derivatives were less active than polymyxin B and one compound contained an Fmoc group in the terminal acyl position.
• a more recent total synthesis of polymyxin B and a few closely related compounds was accomplished only by solid phase peptide synthesis (Ref. 15). Both of these solid phase total synthetic approaches can provide new derivatives of polymyxin but these methods appear limited compared to the methods of the present invention.
• the quantities of antibiotic produced are small and require large amounts of amino acid precursors.
• a primary object of the present invention is to provide potent and effective antibiotics for treating gram-negative bacteria, without limiting use because of toxicity.
• a further object of the present invention is to provide intermediates from polymyxin B or related lipopeptide antibiotics which can be used to prepare new families of antibiotics having potent activity against gram-negative and gram-positive bacteria.
• polymyxin B protected form of polymyxin B is polyanionic, water soluble as a salt, and readily reacts in aqueous solution with a particular deacylase enzyme produced by Actinoplanes utahensis. It has not been found to react with polymyxin deacylase or the other enzymes commonly used to produce the nonapeptides of polymyxin or colistin (Ref. 11).
• the deacylated protected polymyxin peptide designated the protected PBpeptide-3, has three amino acids in the side chain.
• the enzyme from Actinoplanes utahensis can be used as the whole broth from the fermentation, the washed cells, or a water-solubilized preparation.
• the water-solubilized enzyme preparation was obtained by a basic extraction of the washed cells and then the clear extract was adjusted to pH 7-8. This enzyme preparation is the easiest to use, can be freeze-dried to a powder form, and is the most efficient.
• This invention includes methods for preparing four novel protected peptide intermediates from polymyxin B or related lipopeptide antibiotics that are used to prepare new families of anti-biotics which have potent activity against gram-negative, and in some cases gram-positive bacteria.
• Polymyxin B can be isolated from the fermentation of Bacillus polymyxa according to procedures described in Ref. 14. New antibiotics can be prepared from not only polymyxin B but also from related antibiotics such as other polymyxins, colistin, circulin, and octapeptin (EM49, Ref. 13) by these procedures.
• the five basic groups of polymyxin B are protected with a sulfonic acid derivative of 9-fluorenylmethoxycarbonyl, as shown in the following structures of the protecting groups:
• the deacylated protected polymyxin peptide designated the protected PBpeptide-3, has three amino acids in the side chain.
• the enzyme preparation that deacylates the protected polymyxin B can be obtained as a water soluble freeze-dried powder which is relatively stable.
• the preparation of this enzyme is easily accomplished and requires a unique process involving fermentation of Actinoplanes utahensis, separating the cells from the fermentation, washing the cells with water, extracting the cells with a basic buffer at pH 10 for about 20 minutes, adjusting the extract to pH 7-8 and freeze-drying.
• the powdered form of the enzyme resulting from this process is relatively stable and can be readily re-dissolved in water for use. Further purification can be obtained by gel filtration chromatography.
• This enzyme readily deacylates the N-[(2-Sulfo)-9-fluorenlymethoxycarbonyl)] 5 polymyxin B to obtain the protected polymyxin B peptide.
• This soluble enzyme was also effective in current studies for deacylation of N-fluorenlymethoxycarbonyl) amphomycin, laspartomycin, and N-fluorenlymethoxycarbonyl)-A21978C which are not polymyxin.
• polymyxin-type antibiotics such as the polymyxins, octapeptins, colistin, or circulins
• the procedures described above can be used to prepare protected intermediates selected from a group consisting of the following or their corresponding salts:
• the present invention describes semisynthetic antimicrobial cyclopeptides and their methods of preparation from the PBpeptides or other related peptides derived from collistin, circulin or octapeptin.
• This invention describes four different PBpeptide intermediates which can be elaborated further to afford the new antimicrobial derivatives, illustrated in the following embodiment:
• Each of these novel PBpeptide intermediates can be used to make new series of potent antibacterial antibiotics by the general procedures described.
• the desired acids were converted to activated species and then coupled to the protected intermediate PBpeptides and deprotected to give the new antibiotics as represented by compounds 1, 2, 6, 7, 11 and 12 in Table 1.
• the protected intermediate PBpeptides could also be acylated directly with alkyl or aromatic isocyanates or isothiocyanates to give the corresponding ureas and thioureas and the products deprotected to give other new series of antibiotics as represented by compounds 3-5.
• the biological activities of some of the new antibiotics prepared from PBpeptides by these procedures are given in Table 1.
• the HSO 3 Fmoc or other acidic Fmoc protected peptides from colistin (Cpeptides), circulin A (CA peptides), octapeptin B (Obpeptin), octapeptin C (OCpeptides), polymyxin A (PApeptides), and polymyxin D (PDpeptides) can be derivatized to give new antibiotics and new antibiotic prodrugs by the described procedures.
• the new HSO 3 Fmoc derivatives of our new antibiotics are listed in Table 2, and would all be expected to be pro-drugs for their corresponding antibiotics.
• the pro-drug may act as a slow release mechanism for the pharmacokinetics of the antibiotic.
• Polymyxin B sulfate (1.0 g., 0.841 mmol.) was dissolved in a solution of 25 ml., saturated sodium bicarbonate, 25 ml. of water and 25 ml. of tetrahydrofuran.
• a solution of (2-sulfo)-9-fluorenylmethoxy-N-hydroxysuccinimide (2.0 g., 4.8 mmol.) in 25 ml. of tetrahydrofuran was added in several portions over 45 min.
• the reaction mixture was stirred at room temperature over night and diluted with 50 ml. of water, then acidified with 25 ml of 6N hydrochloric acid to give an oily precipitate.
• the deacylase is produced by culturing Actinoplanes utahensis NRRL 12052 under submerged aerobic fermentation conditions. Because single-colony isolates from a lyophile of the culture were heterogeneous for both morphology and enzyme production capability, selections were made to recover a stable, high-producing variant. Initially, multiple fermentations were carried out using inocula prepared from strain 12052. Vegetative growth from the flask yielding the best deacylating activity was plated on a differential agar (CM). Colonies were then selected for further evaluation. Generally, small colonies were better enzyme producers than the large colony types. Isolate No. 18 was selected as a small colony type and shown to be the best deacylase producer of all colonies selected.
• CM differential agar
• CM agar contained corn steep liquor 0.5%, Bacto peptone 0.5%, soluble starch 1.0%, NaCl 0.05%, CaCl 2 .2H 2 O 0.05% and Bacto agar 2.0%.
• the fermentation protocol employed is known (17).
• a high-producing, natural variant was used in this invention.
• the resulting mycelial suspension was transferred into 50 mL of PM3 medium in a 250-mL Erlenmeyer flask.
• This medium contained sucrose 2.0%, peanut meal 1.0%, K 2 HPO 4 0.12%, KH 2 PO 4 0.05% and MgSO 4 .7H 2 O 0.025% in tap water.
• the flask was incubated at a temperature of 30° C. for a period of 60 to 90 hrs.
• the harvest time was determined by an assay which involved HPLC analyses of the deacylation of (2-Sulfo-9-fluorenlymethoxycarbonyl)] 5 polymyxin B by the whole broth at different times during the fermentation.
• Precipitation Method Cells from 450 ml deacylase enzyme were washed 3 ⁇ with water, then brought back to original volume with 0.02 M ammonium phosphate buffer and adjusted to pH 8.0. N-[(2-Sulfo)-9-fluorenlymethoxycarbonyl)] 5 -polymyxin B, 897 mg, was added and the mixture was placed on a shaker at 174 rpm and maintained at 30° C. After five hours the mixture was separated by centrifuging. The clear decant was adjusted to pH 2.3 with 1 N HCl to induce precipitation and allowed to stand at room temperature.
• the precipitate was separated from the mixture, slurried in 80 ml water, adjusted to pH 6.5 to obtain a clear solution, and freeze-dried to obtain 400 mg of tan powder, the semi-purified salt of the protected peptide, N-[(2-Sulfo)-9-fluorenlymethoxycarbonyl] 5 -polymyxin peptide [(NaSO 3 -Fmoc) 5 -PBP-3].
• the cells were extracted with methanol/water to obtain additional material.
• Resin Method Cells from 1 liter deacylase enzyme were washed 3 ⁇ with water, brought back to original volume with 0.02 M ammonium phosphate buffer, and combined with 2.0 g of N-(2-sulfo)-9-fluorenlymethoxycarbonyl)] 5 -polymyxin B. The mixture was placed on a shaker at 175 rpm and maintained at 30° C. for 17 hours. The mixture was then separated by centrifuge and the decant was combined with 20 ml of Amberchrom® CG-161m resin. The resin was washed with 150 ml water, 100 ml 10% CH 3 CN:H 2 O (3 ⁇ ), and 100 ml 20% CH 3 CN:H 2 O (2 ⁇ ).
• the peptide was then eluted 2 ⁇ with 30% CH 3 CN:H 2 O, evaporated to remove CH 3 CN, and freeze-dried to obtain 283 mg powder, the purified peptide.
• the remaining peptide was then eluted 3 ⁇ with 100 ml 50% CH 3 CN:H 2 O, which were combined evaporated and freeze-dried to obtain 460 mg powder, the purified peptide.
• the remaining peptide was extracted from the cells 6 ⁇ with MeOH: H 2 O, 100 ml each. The extracts were combined, brought to four liters with water, adjusted to pH 2.1 with sulfuric acid and combined with Amberchrom® CG-161m resin.
• the resin was rinsed with water and then the peptide was eluted with 100 ml 35% CH 3 CN:H 2 O, which was evaporated and freeze-dried to obtain 94 mg purified peptide. The remaining peptide was eluted with 50% CH 3 CN:H 2 O, evaporated to remove CH 3 CN, and freeze-dried to obtain 539 mg of the purified peptide.
• the [(2-Sulfo)-9-fluorenlymethoxycarbonyl)] 5— polymyxin B peptide[(HSO 3 -Fmoc) 5 -PBP-3)] was isolated from these procedures as tan powder, C 122 H 134 N 16 O 37 S 5 .
• N-phenylthiocarbamyl-(NaSO 3 -Fmoc) 5 -PBP-3 was dissolved in 0.20 mL dimethylformamide (DMF), 0.010 mL piperidine was added and stirred at RT for 60 min.
• the reaction mixture was diluted with 4 mL 0.10M ammonium acetate-0.050M acetic acid (pH 5.05) and 0.006 mL acetic acid and 4 mL MeOH.
• the clear solution was applied to a CM-Sepharose® column (10 ⁇ 20 mm, ca. 2 mL volume) which had been conditioned with 50% MeOH 0.05M in ammonium acetate buffer at pH 5.0.
• the sample-loaded column was rinsed with 4 mL 50% MeOH-0.05M ammonium acetate pH 5.0 then with 4 mL 0.05M ammonium acetate pH 5.0 buffer.
• Product was eluted with 8 mL of 0.27 M sodium sulfate at pH 2.3.
• the product was further purified by application onto a 0.5 g styrene-divinylbenzene cartridge (EnviChrom-P) which was eluted with incrementally increasing concentrations of CH 3 CN 0.05M in pH 2.3 sodium sulfate; product was eluted with 20% CH 3 CN.
• Solvent was removed under vacuum from the product-containing fraction pool which was then desalted in similar fashion as in Example 5, pH adjusted to 6.3 then freeze dried. Yield: 2.7 mg of a white solid, PTC-PBP-3, C 54 H 87 N 17 O 12 S.
• N-phenylthiocarbamyl-(NaSO 3 -Fmoc) 5 -PBP-3 was dissolved in 0.30 mL of anhydrous trifluoroacetic acid (TFA) and heated in a 50° C. water bath for 15 min. TFA was evaporated with a stream of dry nitrogen and the residue was dissolved in 12 mL 0.20M ammonium phosphate at pH7.2 and 6 mL CH 3 CN containing 49 mg triglycine (as an acylating agent scavenger).
• TFA trifluoroacetic acid
• the product solution was first applied to a 0.5 g styrene-divinylbenzene resin cartridge (EnviChrom-P) which was eluted with 10 mL of 40% CH 3 CN 0.04M in ammonium phosphate at pH 7.2.
• Product-containing fractions were pooled ( ⁇ 18 mL), diluted with 12 mL distilled water, then applied to a fresh 0.5 g resin cartridge which was eluted with incrementally increasing concentrations of CH 3 CN about 0.05M in ammonium phosphate at pH 7.2.
• reaction mixture was diluted with 4 mL 0.25M ammonium sulfate at pH2.3 yielding a very milky mixture at pH 3.0 which was extracted with 4 mL ethylacetate; the product-containing aqueous phase was diluted with 4 mL distilled water.
• Product was initially isolated by size exclusion chromatography on a Sephadex® G-25 column (2.5 ⁇ 40 cm) eluted with 0.10M ammonium sulfate at pH 2.3.
• reaction mixture was diluted with 5 mL 0.20M ammonium sulfate at pH2.3 yielding a very milky mixture at pH 2.8 which was extracted with 5 mL ethylacetate.
• Product was initially isolated by size exclusion chromatography on a Sephadex® G-25 column (2.5 ⁇ 40 cm) eluted with 0.10M ammonium sulfate at pH 2.3.
• Product-containing fractions were pooled and further purified on a 0.5 g styrene-divinylbenzene cartridge (EnviChrom-P) by elution with incrementally increasing concentrations of CH 3 CN about 0.10M in ammonium sulfate at pH 2.3; product was eluted with 30% CH 3 CN. Further purification was achieved using size exclusion chromatography on a Sephadex LH-20 column (2.5 ⁇ 40 cm) eluted with 0.026M ammonium acetate/0.053M acetic acid in MeOH. Product-containing fractions were evaporated under vacuum to near dryness, the residue dissolved in 10 mL distilled water, then desalted and freeze dried in similar fashion as in Example 5.
• EnviChrom-P 0.5 g styrene-divinylbenzene cartridge
• Product was further purified on a 0.5 g styrene-divinylbenzene cartridge (EnviChrom-P) using incrementally increasing concentrations of CH 3 CN about 0.05M in sodium sulfate at pH2.3; product was eluted with 25% CH 3 CN. Product-containing fractions were pooled, diluted with an equal volume of distilled water, and product was desalted and freeze dried in similar fashion as in Example 5. Yield: 4.7 mg of a white solid, Compound 7, C 59 H 90 N 16 O 14 .
• Product was isolated on a 0.5 g styrene-divinylbenzene cartridge (EnviChrom-P) using incrementally increasing concentrations of CH 3 CN about 0.05M in sodium sulfate at pH2.3; product was eluted with 25% and 30% CH 3 CN. Product-containing fractions were pooled, solvent removed under vacuum, 2 mL 1.0M ammonium acetate pH 5.0 buffer was added, pH adjusted to 5.0 by addition of ca. 0.6 mL 1.5M NH 4 OH, and then diluted with an equal volume of MeOH. Product was isolated via CM-Sepharose chromatography as in Example 7.
• Example 8 The procedure used in Example 8 to prepare the (NaSO 3 -Fmoc) 4 -PBP-2 from (NaSO 3 -Fmoc) 5 -PBP-3 was repeated starting with (NaSO 3 -Fmoc) 4 -PBP-2, 50 mg, to remove the threonine residue and yielding (NaSO 3 -Fmoc) 4 -PBP-1, 37 mg, as a white powder, with the expected HPLC relative retention time.
• (NaSO 3 -Fmoc) 4 -PBP-2 (5.0 mg) was dissolved in 1.4 mL of 0.02M sodium citrate buffer (pH6.4) and 0.006M in 2-mercaptoethylamine(MEA) hydrochloride.
• the enzyme solution was prepared by dissolving 10 units (0.9 mg) of cathepsin C (dipeptidyl aminopeptidase) (EC 3.4.14.1) in 11.0 mL of the citrate-MEA buffer. Enzyme solution (0.25 mL) was added to the peptide solution which was incubated at about 37° C. for about 24 hours. HPLC analysis indicated about 75% conversion to (NaSO 3 -Fmoc) 3 -PBP.
• the incubated solution was diluted with 11.0 mL 0.4M pH7.2 ammonium phosphate buffer, 3.0 mL distilled water and 0.5 mL CH 3 CN.
• the product was isolated on an ENVI-Chrom-P 0.5 g resin cartridge; product was eluted with 25% CH 3 CN, 0.05M in pH7.2 buffer.
• product-containing fractions (accounting for about 72% of the total product) were pooled, desalted and freeze dried as in Example 8. Yield: 1.0 mg of a pale yellow solid, 87% by HPLC (215 nm area %), with the expected HPLC relative retention time for the (NaSO 3 -Fmoc) 3 -PBP.

Priority Applications (20)

Applications Claiming Priority (1)

Publications (1)

Family

ID=35514884

Family Applications (2)

Family Applications After (1)

Country Status (15)

Cited By (18)

Families Citing this family (22)

Family Cites Families (119)

• 2004
  • 2004-07-01 US US10/881,160 patent/US20060004185A1/en active Pending
• 2005
  • 2005-07-01 WO PCT/US2005/023343 patent/WO2006083317A2/en active Application Filing
  • 2005-07-01 BR BRPI0512941-9A patent/BRPI0512941A/pt not_active IP Right Cessation
  • 2005-07-01 EP EP10184953A patent/EP2332965A1/en not_active Withdrawn
  • 2005-07-01 NZ NZ579261A patent/NZ579261A/en not_active IP Right Cessation
  • 2005-07-01 RU RU2007103811/04A patent/RU2428429C2/ru not_active IP Right Cessation
  • 2005-07-01 JP JP2007519447A patent/JP2008505858A/ja active Pending
  • 2005-07-01 MX MXPA06015239A patent/MXPA06015239A/es not_active Application Discontinuation
  • 2005-07-01 CA CA002571944A patent/CA2571944A1/en not_active Abandoned
  • 2005-07-01 AU AU2005326770A patent/AU2005326770B2/en not_active Ceased
  • 2005-07-01 NZ NZ552730A patent/NZ552730A/en not_active IP Right Cessation
  • 2005-07-01 CN CNA2005800294199A patent/CN101010336A/zh active Pending
  • 2005-07-01 KR KR1020077002649A patent/KR20070047770A/ko not_active Application Discontinuation
  • 2005-07-01 EP EP05856867A patent/EP1761554A2/en not_active Withdrawn
  • 2005-07-01 US US11/630,847 patent/US8889826B2/en not_active Expired - Fee Related
• 2006
  • 2006-12-21 ZA ZA200610818A patent/ZA200610818B/en unknown
  • 2006-12-31 IL IL180458A patent/IL180458A0/en unknown
• 2007
  • 2007-01-30 NO NO20070563A patent/NO20070563L/no not_active Application Discontinuation
• 2011
  • 2011-04-15 RU RU2011115077/04A patent/RU2011115077A/ru not_active Application Discontinuation
  • 2011-08-01 JP JP2011168634A patent/JP2011256189A/ja active Pending

Also Published As

Similar Documents

Legal Events