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WO2007080108A1 - Composition ascorbic acid and a metal ion for selective killing of pathological mammalian cells - Google Patents

Composition ascorbic acid and a metal ion for selective killing of pathological mammalian cells Download PDF

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Publication number
WO2007080108A1
WO2007080108A1 PCT/EP2007/000164 EP2007000164W WO2007080108A1 WO 2007080108 A1 WO2007080108 A1 WO 2007080108A1 EP 2007000164 W EP2007000164 W EP 2007000164W WO 2007080108 A1 WO2007080108 A1 WO 2007080108A1
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WO
WIPO (PCT)
Prior art keywords
metal ion
ascorbic acid
cells
mammalian cells
pathologically altered
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.)
Ceased
Application number
PCT/EP2007/000164
Other languages
French (fr)
Inventor
Thomas Lisowsky
Karlheinz Esser
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Multibind Biotec GmbH
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Multibind Biotec GmbH
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Multibind Biotec GmbH filed Critical Multibind Biotec GmbH
Publication of WO2007080108A1 publication Critical patent/WO2007080108A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00Medicinal preparations containing inorganic active ingredients
    • A61K33/24Heavy metals; Compounds thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00Medicinal preparations containing inorganic active ingredients
    • A61K33/24Heavy metals; Compounds thereof
    • A61K33/26Iron; Compounds thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365Lactones
    • A61K31/375Ascorbic acid, i.e. vitamin C; Salts thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00Medicinal preparations containing inorganic active ingredients
    • A61K33/24Heavy metals; Compounds thereof
    • A61K33/30Zinc; Compounds thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00Medicinal preparations containing inorganic active ingredients
    • A61K33/24Heavy metals; Compounds thereof
    • A61K33/34Copper; Compounds thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents

Definitions

  • the invention concerns a composition for selective killing and destruction of pathological mammalian cells and uses of this composition.
  • composition comprising a synergistic mixture of a) ascorbic acid and b) at least one metal ion.
  • An additional enhancement of the effect of the two components, i.e. vitamin c or derivates thereof and at least one metal ion, can be achieved by the optional addition of factors that promote the up-take of metal ions into pathologically altered cells.
  • factors are for example natural transferrin receptors or the human growth factor Augmenter of Liver Regeneration protein (Alrp) [Lange et al.
  • One embodiment of the present invention comprises solutions containing a synergistic mixture of a) at least ascorbic acid or derivates thereof and b) at least one metal ion and c) optionally selected substances that support the up-take of metal ions.
  • Another purpose of the present invention is the use of the composition according to the invention for selective killing and destruction of pathologically altered mammalian cells.
  • ascorbic acid or derivates thereof with at least one metal ion results in a synergistic effect and in rapid, efficient and selective killing of pathologically altered mammalian cells.
  • the components are provided in appropriate concentrations.
  • composition according to the invention shows selective killing and destruction of pathological cells in the pH range of 1 to 6,5 with an essentially comparable efficiency.
  • Adjustment and consideration of correct pH values for the solutions according to the invention is therefore preferred.
  • ascorbic acid, their related salts or acidic derivates are used as well as at least one compound selected from the group of the water-soluble forms having the characteristics of an antioxidant.
  • the derivates of ascorbic acid or their respective salts or acidic derivates are one or several compounds and/or their related salts selected from the group of the highly water-soluble compounds. They are provided in concentrations of 0,1 mM to 50 mM in relation to the total volume of the solution, preferably, in concentrations of 0.1 mM to 5 mM.
  • the metal ions according to the invention are di- and/or tri- valent ions of metals found in the 4th period and/or subgroup I, II and VIII (IB, HB and VIIIB) of the periodic table of the elements. They are provided as salts in combination with their organic and/or inorganic acids and bases. According to the Invention, one or several compounds selected from sub-group VIII, especially iron, cobalt, nickel, copper or zinc are preferred. They are provided in concentrations of 0,01 itiM to 5 inM, in relation to the total volume of the solution, preferably in concentrations of 0.1 mM to 2 inM.
  • composition according to the invention can contain additional common inert adjuvants and additives like for example suitable buffer substances for defining a specific pH value, like Tris (Tris (hydroxymethyl) -aminomethan) , MES (2 (Morpholino)ethansulfonic acid), HEPES (2- [4- (2- Hydroxyethyl) -1-piperazinyl] -ethansulfonic acid, MOPS (3-(N- Morpholino) propansulfonic acid), carbonate and derivates of succinic acid.
  • suitable buffer substances for defining a specific pH value
  • suitable buffer substances like Tris (Tris (hydroxymethyl) -aminomethan) , MES (2 (Morpholino)ethansulfonic acid), HEPES (2- [4- (2- Hydroxyethyl) -1-piperazinyl] -ethansulfonic acid, MOPS (3-(N- Morpholino) propansulfonic acid), carbonate
  • the application in general takes place through intra-venous injection via the blood or by direct application into the pathologically altered cells.
  • Fig. 1 shows the principle for selective killing of pathologically altered cells by the composition according to the invention.
  • the up to thousand-fold accumulation of metal ions in pathologically altered cells results in sensitisation towards ascorbic acid and derivates thereof.
  • Combination of the application of ascorbic acid and derivates thereof with at least one additional metal ion and optionally with substances that support the up-take of metal ions results in selective killing of the pathologically altered cells.
  • Advantageous for this effect is the synergistic mixture of the components and preferably the compliance of well defined concentrations that have no deleterious effect on normal cells but that induce apoptosis for pathological cells .
  • Table 1 shows a preferred basic composition and a preferred mixture for a two component system made of ascorbic acid and/or derivates thereof and metal ions.
  • composition of the solutions pH range 1.0 to 6.5 ascorbic acid 0.1 mM to 5 mM metal ions: 0 01 mM to 2 mM proteins to enhance the up-take of metal ions: 1 mg to lOOmg per 1 kg body weight
  • Table 1 Preferred compositions of a solution comprising ascorbic acid and/or derivates thereof, metal ions and optionally different substances that support up-take of metal ions into pathologically altered mammalian cells. List with explanation of the abbreviations in the figures
  • Alrp Augmenter of Liver Regeneration protein
  • Apoptosis programmed cell death

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Inorganic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention concerns a composition comprising vitamin C and/or derivates thereof and at least one metal ion for selective and efficient killing and/or destrcution of pathologically altered mammalian cells. The solutions according to the invention are applied intra-venously via blood or by direct injection into the pathologically altered united cell structure. Thereby these cells are efficiently and selectively killed and/or destroyed. Additional enhancement of this effect can be achieved by the optional application of substances that support up-take of metal ions into pathologically altered cells.

Description

COMPOSITIONASCORBIC ACID AND A METAL ION FOR SELECTIVE KILLING OF PATHOLOGICAL MAMMALIAN CELLS
Field of the invention
The invention concerns a composition for selective killing and destruction of pathological mammalian cells and uses of this composition.
Background of the invention
The latest discovery of the selective action of ascorbic acid on pathologically altered mammalian cells offers new chances for therapies [Chen et al. (2005)].
Pathologically altered mammalian cells play a special role in wound healing, infections and tumorigenesis. For a long time manifold therapeutical approaches are aiming for the selective killing of such cells.
Another approach for the improvement of such therapies offers the finding that pathological mammalian cells that have an abnormally high and degenerated division rate also accumulate iron up to a thousand-fold higher than normal, healthy cells [Weinberg (1999) ] .
Prior art
According to these findings the prior art for possible therapies is based on the reduction of the iron concentration in pathologically altered cells to a normal level to achieve curing or to support the therapy [Abeysinghe et al . (2001)]. Therefore in latest studies for the action of ascorbic acid on tumor cells included also tests for combinations with chelating agents for metal ions. In these studies no significant synergistic effect was found for the action of ascorbic acid in combination with metal iron chelators.
Therefore for practical applications there is the necessity for new improved protocols, agents, methods and compositions for efficient and particularly gentle selective killing and destruction of pathologically altered cells.
Summary of the invention
It is therefore the objective of the present invention to overcome the current limitations and disadvantages of the prior art and to develop a new composition that does allow selective and efficient killing and destruction of pathological mammalian cells under conditions that are preferably not harmful for normal cells.
This objective is achieved by a composition comprising a synergistic mixture of a) ascorbic acid and b) at least one metal ion.
In contrast to prior art a new system was developed by combining ascorbic acid and at least one metal ion for the selective killing and destruction of pathologically altered mammalian cells. Thereby the synergistic action of the at least two substances results in a higher efficiency, a more rapid action and the use of lower concentrations of all components . The new more sensitive effect of killing is achieved by additional accumulation of metal ions in pathologically altered cells. These metal ions in combination with ascorbic acid result in enhanced formation of radicals and the generation of hydrogen peroxide. The synergistic action of all components results in selective damage of pathologically- altered cells with damages at membranes and internal DNA and RNA molecules, leading to induction of programmed cell death, the so-called apoptosis.
The use of natural biomolecules that are naturally present in all living cells thereby minimizes any possible side effects.
An additional enhancement of the effect of the two components, i.e. vitamin c or derivates thereof and at least one metal ion, can be achieved by the optional addition of factors that promote the up-take of metal ions into pathologically altered cells. Such factors are for example natural transferrin receptors or the human growth factor Augmenter of Liver Regeneration protein (Alrp) [Lange et al.
(2001) ] . Such factors can support a higher up-take of metal ions into pathologically altered cells.
Medical and commercial interest in such solutions is notedly high.
One embodiment of the present invention comprises solutions containing a synergistic mixture of a) at least ascorbic acid or derivates thereof and b) at least one metal ion and c) optionally selected substances that support the up-take of metal ions. Another purpose of the present invention is the use of the composition according to the invention for selective killing and destruction of pathologically altered mammalian cells.
The combination of ascorbic acid or derivates thereof with at least one metal ion, preferably in an appropriate mixture, results in a synergistic effect and in rapid, efficient and selective killing of pathologically altered mammalian cells. Preferably, in the composition according to the invention the components are provided in appropriate concentrations.
Especially surprising and new is the finding that the composition according to the invention shows selective killing and destruction of pathological cells in the pH range of 1 to 6,5 with an essentially comparable efficiency.
Adjustment and consideration of correct pH values for the solutions according to the invention is therefore preferred.
According to the invention, it is particularly preferred that ascorbic acid, their related salts or acidic derivates are used as well as at least one compound selected from the group of the water-soluble forms having the characteristics of an antioxidant. According to the invention, the derivates of ascorbic acid or their respective salts or acidic derivates are one or several compounds and/or their related salts selected from the group of the highly water-soluble compounds. They are provided in concentrations of 0,1 mM to 50 mM in relation to the total volume of the solution, preferably, in concentrations of 0.1 mM to 5 mM.
The metal ions according to the invention are di- and/or tri- valent ions of metals found in the 4th period and/or subgroup I, II and VIII (IB, HB and VIIIB) of the periodic table of the elements. They are provided as salts in combination with their organic and/or inorganic acids and bases. According to the Invention, one or several compounds selected from sub-group VIII, especially iron, cobalt, nickel, copper or zinc are preferred. They are provided in concentrations of 0,01 itiM to 5 inM, in relation to the total volume of the solution, preferably in concentrations of 0.1 mM to 2 inM.
The composition according to the invention can contain additional common inert adjuvants and additives like for example suitable buffer substances for defining a specific pH value, like Tris (Tris (hydroxymethyl) -aminomethan) , MES (2 (Morpholino)ethansulfonic acid), HEPES (2- [4- (2- Hydroxyethyl) -1-piperazinyl] -ethansulfonic acid, MOPS (3-(N- Morpholino) propansulfonic acid), carbonate and derivates of succinic acid. The buffer systems are provided in concentrations of i mM to 500 mM in relation to the total volume of the solution.
In principle thereby all kind of pathologically altered cells with an especially high level of iron, enhanced iron up-take and abnormally high cell division rates can be killed selectively, efficiently and in a way that is not harmful and gentle for normal cells
The application in general takes place through intra-venous injection via the blood or by direct application into the pathologically altered cells.
Various and preferred embodiments of the invention
The invention is illustrated by nonrestrictive figures, examples and tables shown in the following part: In that way Fig. 1 shows the principle for selective killing of pathologically altered cells by the composition according to the invention. The up to thousand-fold accumulation of metal ions in pathologically altered cells results in sensitisation towards ascorbic acid and derivates thereof.
Combination of the application of ascorbic acid and derivates thereof with at least one additional metal ion and optionally with substances that support the up-take of metal ions results in selective killing of the pathologically altered cells. Advantageous for this effect is the synergistic mixture of the components and preferably the compliance of well defined concentrations that have no deleterious effect on normal cells but that induce apoptosis for pathological cells .
Table 1 shows a preferred basic composition and a preferred mixture for a two component system made of ascorbic acid and/or derivates thereof and metal ions.
composition of the solutions pH range 1.0 to 6.5 ascorbic acid 0.1 mM to 5 mM metal ions: 0 01 mM to 2 mM proteins to enhance the up-take of metal ions: 1 mg to lOOmg per 1 kg body weight
Table 1: Preferred compositions of a solution comprising ascorbic acid and/or derivates thereof, metal ions and optionally different substances that support up-take of metal ions into pathologically altered mammalian cells. List with explanation of the abbreviations in the figures
Alrp: Augmenter of Liver Regeneration protein Apoptosis: programmed cell death
References
Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, Shacter E & Levine M (2005): Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci U.S.A. 102 ( 38 ): 13604-13609
Weinberg ED (1999): Iron Loading and Disease Surveillance. Emerging Infectious Diseases 5(3): 346 -352
Abeysinghe RD, Greene BT, Haynes R., Willingham M. C, Turner J., Planalp R. P., Brechbiel M.W. , Torti F. M. & Torti S. V. (2001) : p53-independent apoptosis mediated by tachpyridine, an anti-cancer iron chelator. Carcinogenesis. 22(10): 1607- 1614
Lange H, Lisowsky T, Gerber J, Mϋhlenhoff U, Kispal G, LiIl R (2001) : Essential function of the mitochondrial sulfhydryl oxidase Ervlp/Alrp in the maturation of cytosolic Fe/S proteins. EMBO 2 (8):715-720

Claims

Claims
1. Composition comprising a synergistic mixture of a) ascorbic acid and b) at least one metal ion.
2. Composition according to claim 1, wherein the metal ion is selected from the metals of the 4th period and sub-groups
I, II, or VIII of the periodic table of the elements.
3. Composition according to claim 1 or 2, wherein the metal ion is provided as a salt of the respective acid or base.
4. Composition according to at least one of the claims 1 to
3, wherein the ascorbic acid or derivates thereof is/are provided in concentrations of 0.1 mM to 50 mM.
5. Composition according to at least one of the claims 1 to
4, wherein the metal ion is provided in concentrations of 0.01 mM to 5 mM.
6. Composition according to at least one of the claims 1 to 5, wherein additional substances are added that enhance the cellular up-take of metal ions.
7. Use of the composition according to at least one of the claims 1 to 6 for selective killing and/or destruction of pathologically altered mammalian cells.
PCT/EP2007/000164 2006-01-10 2007-01-10 Composition ascorbic acid and a metal ion for selective killing of pathological mammalian cells Ceased WO2007080108A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102006001213A DE102006001213A1 (en) 2006-01-10 2006-01-10 Synergistic solution of ascorbic acid or its derivatives and metal ions efficient, selective killing of pathological mammalian cells
DE102006001213.5 2006-01-10

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WO2007080108A1 true WO2007080108A1 (en) 2007-07-19

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9795133B2 (en) 2011-01-26 2017-10-24 Multibind Biotec Gmbh Synergistic disinfecting compositions with essential oils
US10980239B2 (en) 2005-04-30 2021-04-20 Multibind Biotec Gmbh Decontamination solution and its use for denaturation, modification, degradation, solubilisation and removal of proteins, nucleic acid molecules and microorganisms

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2041M (en) * 1962-04-17 1963-09-30 Andre Anselme Marie Therapeutic composition with cytotoxic properties based on ascorbic acid and metal ions.
WO1999051585A1 (en) * 1998-04-07 1999-10-14 Even Marstrand Composition for the treatment of cancer and colitis ulcerosa as well as the use of a titanium complex and a zinc salt for the preparation of the composition
WO2001024803A2 (en) * 1999-10-04 2001-04-12 John Carter Pharmaceutical compositions containing copper, salicylic acid and vitamines c
WO2001082938A2 (en) * 2000-04-28 2001-11-08 Ocean Nutrition Canada Limited Compositions useful in the treatment of diseases of connective tissues comprising a ferrous ion and an ascorbate

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2041M (en) * 1962-04-17 1963-09-30 Andre Anselme Marie Therapeutic composition with cytotoxic properties based on ascorbic acid and metal ions.
WO1999051585A1 (en) * 1998-04-07 1999-10-14 Even Marstrand Composition for the treatment of cancer and colitis ulcerosa as well as the use of a titanium complex and a zinc salt for the preparation of the composition
WO2001024803A2 (en) * 1999-10-04 2001-04-12 John Carter Pharmaceutical compositions containing copper, salicylic acid and vitamines c
WO2001082938A2 (en) * 2000-04-28 2001-11-08 Ocean Nutrition Canada Limited Compositions useful in the treatment of diseases of connective tissues comprising a ferrous ion and an ascorbate

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
BRAM S ET AL: "VITAMIN C PREFERENTIAL TOXICITY FOR MALIGNANT MELANOMA CELLS", NATURE, MACMILLAN JOURNALS, LONDON, GB, vol. 284, no. 5757, 1980, pages 629 - 631, XP008079336, ISSN: 0028-0836 *
GONZÁLEZ MICHAEL J ET AL: "Inhibition of human breast carcinoma cell proliferation by ascorbate and copper.", PUERTO RICO HEALTH SCIENCES JOURNAL MAR 2002, vol. 21, no. 1, March 2002 (2002-03-01), pages 21 - 23, XP008079367, ISSN: 0738-0658 *
LEUNG P Y ET AL: "ASCORBIC ACID WITH CUPRIC IONS AS A CHEMOTHERAPY FOR HUMAN LUNG TUMOR XENOGRAFTS IMPLANTED BENEATH THE RENAL CAPSULE OF IMMUNOCOMPETENT MICE", IN VIVO - INTERNATIONAL JOURNAL OF IN VIVO RESEARCH, XX, GB, vol. 6, no. 1, 1992, pages 33 - 40, XP008079419, ISSN: 0258-851X *
PEREZ YARITZA ET AL: "ASCORBIC ACID AND COPPER: EFFECTIVE CYTOTOXIC AGENTS AGAINST METASTATIC BREAST CARCINOMA (BRCA)CELLS", PROCEEDINGS OF THE ANNUAL MEETING OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH, NEW YORK, NY, US, vol. 42, March 2001 (2001-03-01), pages 215, XP001248648, ISSN: 0197-016X *
SATOH KAZUE ET AL: "EFFECT OF COPPER AND IRON IONS ON CYTOTOXICITY INDUCED BY ASCORBATE, GALLATE AND CAFFEATE", ANTICANCER RESEARCH, HELENIC ANTICANCER INSTITUTE, ATHENS,, GR, vol. 17, no. 3C, 1997, pages 2181 - 2184, XP008079363, ISSN: 0250-7005 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10980239B2 (en) 2005-04-30 2021-04-20 Multibind Biotec Gmbh Decontamination solution and its use for denaturation, modification, degradation, solubilisation and removal of proteins, nucleic acid molecules and microorganisms
US9795133B2 (en) 2011-01-26 2017-10-24 Multibind Biotec Gmbh Synergistic disinfecting compositions with essential oils

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