CN117202918A - Use of methane-oxidizing mycotins for the treatment of iron-related disorders - Google Patents

Abstract

The invention relates to a method for preparing Fe for medicine ³⁺ Reduction of ions to Fe ²⁺ Ionic methane-oxidizing bacteria and pharmaceutical compositions comprising same and in vitro Fe-oxidation of bacteria ³⁺ Reduction of ions to Fe ²⁺ Ion method.

Description

Use of methane-oxidizing mycotins for the treatment of iron-related disorders

Technical Field

The invention relates to a method for preparing Fe for medicine (medicine) ³⁺ Reduction of ions to Fe ²⁺ Ionic methane-oxidizing bacteria (methane-oxidizing bacteria) and pharmaceutical compositions comprising said methane-oxidizing bacteria and Fe ³⁺ Reduction of ions to Fe ²⁺ Is a method of (2).

Background

Iron overload can occur in patients with hereditary hemochromatosis, thalassemia, sickle cell disease, aplastic anemia, myelodysplasia and other diseases. Hereditary hemochromatosis is caused by mutations in genes encoding proteins involved in limiting systemic iron uptake. About 10% of the population is heterozygote carriers, 0.3-0.5% homozygous. Currently, these diseases caused by imbalance of iron intake are treated by periodic bleeding (up to 500 ml/week |) or iron chelators such as deferiprox, deferasirox (Exjade, novartis Net Sales2019:995mio$, https:// www.novartis.com/Investors/Financial-Data/Product-Sales) and deferoxamine (Desferal). These chelators have undesirable side effects and are not as effective as bleeding. However, they are only able to partially dissolve the iron deposit.

In addition, brain iron accumulation is associated with several neurodegenerative diseases, such as Alzheimer's disease, parkinson's disease, dementia, huntington's disease (Liu et al, front neurosci.2018;12:632; agraval et al, free Liotic and Medicine 2018,120,317-329; moon et al, J Alzheimer's disease 2016, 51 (3), 737-45) and multiple sclerosis (Stephenson et al, nature Reviews Neurology,2014, volume 10, 459-468).

In particular, iron and iron accumulation play a role in aging (senescence) (Masalman et al, redox Biol,2018, 14:100-115), aging (aging) (Timmers et al, NATURE COMMUNICATIONS | (2020), 11,3570), iron Cell death (Li et al, cell treatment & Disease,11, article number: 88), alcoholic liver Disease (Kowdley, gastroenterol Hepatol (NY), 2016,12 (11): 695-698), or amyotrophic lateral sclerosis (Gajoowiak et al, postepy hig Med Dosw (online), 2016Jun 30;70 (0): 709-21).

Thus, it may be very beneficial to identify compounds that can aid in the consumption of iron, in particular, the dissolution of iron deposits.

Brief description of the invention

The invention relates to a method for preparing Fe for medicine ³⁺ Reduction of ions to Fe ²⁺ Ionic methane-oxidizing bacteria.

Furthermore, the present invention relates to a pharmaceutical composition comprising methane-oxidizing bacteria.

Furthermore, the present invention relates to the in vitro purification of Fe ³⁺ Reduction to Fe ²⁺ Is a method of (2).

As shown in the examples, it has surprisingly been found that certain methane-oxidizing bacteria are capable of complexing Fe ³⁺ Ions and reduce them to Fe ²⁺ . Most of the excess iron is usually replaced by ferritin as Fe ³⁺ Stored in animals, plants and bacteria. Thus, excessive iron Fe ³⁺ Ions can be removed by the methane-oxidizing bacteria found in the present invention. Thus, the methane-oxidizing mycotins of the present invention are useful for treating diseases caused by the accumulation of iron ions in the body.

Brief description of the drawings

Fig. 1: binding and reduction of ferric iron to ferrous iron by MB-SB2

A. 50nmol ml of isolated ^-1 SB2-MB followed by addition of 5nmol FeCl ₃ Ultraviolet-visible absorption spectrum of (c). B. The absorbance of oxazolone (. Smallcircle.) and imidazolone (. DELTA.) groups at 336 and 387nm, respectively, was taken as FeCl ₃ Function of the molar ratio to MB-SB2.

FIG. 2 iron reductase Activity of MB-SB2 but not MB-OB3b

Containing 1mM of ferrioxazine plus 10mM of FeCl ₃ (- - -), 1mM iron oxazine plus 23.4. Mu.M MB-SB (- -), 1mM iron oxazine plus 10mM FeCl ₃ And a reaction mixture of 5.8 (-), 11.6 (-), 17.4 (-) or 23.4 (-) mu M MB-SB2 (A) or MB-OB3B (B) at 562nmChanges in luminosity. C. 4 hours after the addition of MB-SB2, aqueous 4M FeCl ₃ Solution (a) and 4M FeCl ₃ The solution was added with 20mM MB-SB2.

Fig. 3: bile iron excretion by MB-SB2 rather than MB-OB3b

A. Bile iron excretion. Upon liver perfusion, MB-SB2 brings iron to bile, whereas MB-OB3b does not. B. Fecal iron excretion. Intraperitoneal injection of MB-SB2 LPP Atp7b ^-/- After rats, iron was excreted in the feces, but MB-OB3b was not excreted after injection. The dashed line represents the average fecal iron excretion from untreated rats.

Fig. 4: coupling of Water Oxidation with reduction of Fe (III) by MB-SB2

Adding 20mM FeCl ₃ After that, it contains 97% H ₂ ¹⁸ Mass spectrum of the headspace of the reaction mixture of 2mM MB-SB2 in O.

Fig. 5: control Huh7 cells (human hepatoma cell line) were preloaded with 50. Mu.M FAC for 24 hours. Cells were then treated with 0.5mM MB-SB2, 0.5mM MB-OB3b and 0.5mM DFO for 24 hours. In contrast to OB3B and DFO, MB-SB2 significantly reduced the cellular iron concentration in iron preloaded Huh7 cells to untreated levels. Statistical analysis by one-way analysis of variance(N＝4)Is carried out.

FAC: ferric ammonium citrate

UT: untreated Huh7 cells

Control: cells treated with 50. Mu.M FAC for 24 hours

DFO: deferoxamine.

Detailed Description

The solutions of the invention are described below, illustrated in the accompanying examples, illustrated in the accompanying drawings and reflected in the claims.

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The term "and/or" as used herein includes the meaning of "and", "or" and "all or any other combination of the elements connected by the term.

Throughout the specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. The term "comprising" as used herein may be replaced with the term "containing" or "including" or sometimes with the term "having" as used herein. As used herein, "consisting of … …" excludes any elements, steps, or components not specified.

It is to be understood that this invention is not limited to the particular methodology, protocols, materials, reagents, materials, etc., described herein, as such may vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

The invention relates to a method for preparing Fe for medicine ³⁺ Reduction of ions to Fe ²⁺ Ionic methane-oxidizing bacteria.

The term "methane-oxidizing agent" as used herein includes in particular modified peptides characterized by the presence of one oxazolone ring and a second oxazolone, imidazolone or pyrazinedione ring. The two loops are separated by 2-5 amino acid residues. Each ring has an adjacent thioamide group.

Preferably, the methane-oxidizing mycomycin is MB-SB2 and comprises a primary structure according to formula (I). More preferably, the methane-oxidizing mycomycin has a primary structure according to formula (I) and is MB-SB2.

When complexing Fe ²⁺ Or Fe (Fe) ³⁺ When the resulting methane-oxidizing rhzomorph complex is expected to have the structural formula (II). Thus, fe is as follows ³⁺ Reduction to Fe ²⁺ Is envisaged to include intermediate structures according to formula (II).

As used herein, the terms "complex" and "bind" may be used with respect to one anotherThe use of the term "binding" of iron, i.e. for example, methane-oxidizing bacteria, is to be understood as "complexing" iron with methane-oxidizing bacteria and vice versa. The term "complexation" generally refers to the formation of a complex consisting of a central ion and a surrounding array of molecules called ligands or complexing agents. For the purposes of the present invention, the central ion is iron (i.e., fe ²⁺ Or Fe (Fe) ³⁺ ) The ligand is methane-oxidizing mycotin. Methane-oxidizing bacteria will typically complex with an iron ion to form methane-oxidizing bacteria-iron complexes, respectively.

In excess of Fe ³⁺ Preferably from 0.1 to 200, more preferably from 0.5 to 5, most preferably from 0.7 to 3, particularly preferably 1 Fe per minute per methane-oxidizing agent reduction of methane-oxidizing agent ³⁺ Will Fe at a rate of (2) ³⁺ Reduction to Fe ²⁺ 。

Preferably, the reduction is carried out catalytically, using methane-oxidizing mycomycin as catalyst. In the present invention, the term "catalytic" is defined as a reaction in which the "catalyst" molecule increases the reaction rate of the reaction by forming an intermediate, desirably in a manner that achieves a technically useful overall reaction rate. In the present invention, the "catalyst" is regenerated after formation of the intermediate, releasing the regenerated catalyst and the desired reaction product of formula (III).

The methane-oxidizing bacteria of the present invention can utilize various electron donors for Fe ³⁺ And (5) reduction. Preferably, the reducing agent is H ₂ O, molecular oxygen is generated during the reduction process.

It is conceivable that the reduction is preferably performed according to the following equation:

4FeCl ₃ +methane-oxidizing rhzomorph +2H ₂ O _→ 3Fe ²⁺ +Fe (II) -methane-oxidizing bacteria element +12Cl- +4H ⁺ +O ₂ 。

Those skilled in the art will readily appreciate that the methane-oxidizing agent-iron complex is typically formed after administration of the methane-oxidizing agent to a subject, at which time the methane-oxidizing agent complex may deplete (excess) iron in the subject.

The term "methane-oxidizing agent" includes naturally occurring methane-oxidizing agents, and functional variants, fragments, and derivatives thereof, that retain complex iron (i.e., fe ²⁺ And Fe (Fe) ³⁺ ) And preferably binds Fe with a binding affinity comparable to or even higher than that of naturally occurring methane-oxidizing bacteria ³⁺ 。

The term "methanotrophic hormone variant" refers to a methanotrophic hormone of the general formula of the "parent" methanotrophic hormone, but containing at least one amino acid substitution, deletion or insertion as compared to the parent methanotrophic hormone, provided that the variant retains the desired iron binding affinity and/or biological activity described herein.

A "methane-oxidizing rhzomorph derivative" is a chemically modified methane-oxidizing rhzomorph. Generally, all types of modifications are included in the present invention, provided they do not negate the beneficial effects of methane-oxidizing mycotoxins. That is, the methane-oxidizing rhzomorph derivatives preferably retain the iron binding affinity and/or biological activity of the methane-oxidizing rhzomorph from which they are derived. The methane-oxidizing rhzomorph derivatives also include stabilized methane-oxidizing rhzomorphs as described below.

Possible chemical modifications in the context of the present invention include acylation, acetylation or amidation of amino acid residues. Other suitable modifications include, for example, extending amino groups with polymer chains of different lengths (e.g., XTEN technology or) N-glycosylation, O-glycosylation and carbohydrates such as hydroxyethyl starch (e.g.)>) Or polysialic acid (e.g.)>Technology) is provided. Chemical modifications such as alkylation (e.g., methylation, propylation, butylation), arylation and etherification are possible and are also contemplated. Further chemical modifications contemplated herein are ubiquitination, binding to therapeutic or diagnostic agents, labeling (e.g., with radionuclides or various enzymes), and insertion or substitution by chemically synthesizing unnatural amino acids.

Other possible modifications may include removal of the sulfate group and/or replacement of the oxazolonyl group with a more stable imidazolone or pyrazinedionyl group. Addition and/or deletion of genes in the operon of class II methane-oxidizing bacteria to class I or vice versa will result in a change in loop type (note: class I and class II methane-oxidizing bacteria are described in Semrau et al 2020.FEMS Microbiol Lett.367:fn045). Substitution of oxazolone groups with imidazolone or pyrazindione groups should increase the stability of the methane-oxidizing bacteria to the extent possible for oral administration.

For the purposes of the present invention, the methanotrophic mycotins as defined above also include pharmaceutically acceptable salts thereof. The phrase "pharmaceutically acceptable salts" as used herein refers to those salts of methane-oxidizing bacteria that are safe and effective for treatment. Pharmaceutically acceptable salts include salts formed with anions, such as those derived from hydrochloric acid, phosphoric acid, acetic acid, oxalic acid, tartaric acid, choline, and the like, and salts formed with cations, such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxide, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

As previously mentioned, the methane-oxidizing agent fragments, variants, and derivatives preferably retain the advantageous capabilities of methane-oxidizing agents as evaluated in the appended examples.

The methane-oxidizing mycotoxins may be derived from Methylocystis strain SB2 (Methylocystis sp.strain SB 2).

As described above and shown in example 2, the methane-oxidizing bacteria according to the present invention can be produced by adding an excess of iron Fe ³⁺ Reduction of ions to Fe ²⁺ And complex them into the corresponding Fe ²⁺ Ion form to remove excessive iron Fe ³⁺ Ions. Thus, the methane-oxidizing mycotins of the present invention are useful in the treatment of diseases caused by the accumulation of iron ions and the formation of iron precipitates in the body. As also mentioned in the introduction, the following diseases are associated with iron accumulation, and thus their treatment benefitsIn an agent that aids in the consumption of iron deposition, such as methane-oxidizing mycotins of the present invention.

Furthermore, the present invention relates to methanotrophic mycoses for use in the treatment of iron overload disorders, neurodegenerative diseases, iron overload caused by red blood cell transfusion, ageing, aging, iron cell death (ferroptotic cell death), alcoholic liver disease or amyotrophic lateral sclerosis.

Iron overload disorders may be caused by a disease selected from the group consisting of hereditary hemochromatosis, thalassemia, sickle cell disease, aplastic anemia, and myelodysplasia.

The neurodegenerative disease may be selected from the group consisting of Alzheimer's disease, parkinson's disease, dementia, huntington's disease and multiple sclerosis.

The invention and its advantages will be better understood from the following examples, which are for illustrative purposes only. These examples are not intended to limit the scope of the invention in any way.

The invention also relates to a pharmaceutical composition comprising a methane-oxidizing agent as described above for use in medicine, in particular for use in the above indications.

As mentioned above, pharmaceutical compositions comprising methane-oxidizing bacteria are also contemplated herein. Accordingly, other aspects of the invention include pharmaceutical compositions comprising a methane-oxidizing agent as described herein and the use of the methane-oxidizing agent for the manufacture of a pharmaceutical composition. The term "pharmaceutical composition" particularly refers to a composition suitable for administration to a human. However, compositions suitable for administration to non-human animals are also contemplated herein.

The pharmaceutical compositions and components thereof (i.e., the active ingredient and optional excipients or carriers) are preferably pharmaceutically acceptable, i.e., capable of eliciting a desired therapeutic effect without eliciting an undesired or at least acceptable local or systemic effect. The pharmaceutically acceptable compositions of the invention may be in particular sterile and/or pharmaceutically inert. In particular, the term "pharmaceutically acceptable" may refer to use in animals, more particularly in humans, approved by a regulatory agency or other recognized pharmacopoeia.

The methane-oxidizing bacteria described herein are preferably present in the pharmaceutical composition in a therapeutically effective amount.

"therapeutically effective amount" refers to the amount of methane-oxidizing bacteria that results in the desired therapeutic effect. The exact dosage will depend on the purpose of the treatment and will be determined by one skilled in the art using known techniques. Therapeutic efficacy and toxicity can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED ₅₀ (therapeutically effective dose in 50% of population) and LD ₅₀ (dose lethal to 50% of the population). The dose ratio between therapeutic effect and toxic effect is the therapeutic index, which can be expressed as the ratio ED ₅₀ /LD ₅₀ . Pharmaceutical compositions exhibiting a large therapeutic index are generally preferred.

It is envisaged that the pharmaceutical composition comprises a methanotrophic hormone as described herein, in particular a methanotrophic hormone in stable form, and preferably a therapeutically effective amount of a methanotrophic hormone, optionally together with one or more carriers, excipients and/or additional active agents.

"excipient" includes fillers, binders, disintegrants, coatings, adsorbents, anti-adherent agents, glidants, preservatives, antioxidants, flavoring agents, colorants, sweeteners, solvents, co-solvents, buffers, chelating agents, viscosity-imparting agents, surfactants, diluents, wetting agents, carriers, diluents, preservatives, emulsifiers, stabilizers, and tonicity adjusting agents. Exemplary suitable carriers for pharmaceutical compositions of the invention include saline, buffered saline, dextrose, and water.

The pharmaceutical compositions of the invention may be formulated in various forms, for example solid, liquid, gaseous or lyophilized forms, and may be in particular in the form of ointments, creams, transdermal patches, gels, powders, tablets, solutions, aerosols, granules, pills, suspensions, emulsions, capsules, syrups, liquids, elixirs, extracts, tinctures or fluid extracts (fluid extracts), or in a form particularly suitable for the desired method of administration. Methods for preparing medicaments known per se are described in Forth, henschler, rummel (1996) Allgemeine und spezielle Pharmakologie und Toxikologie, urban & Fischer.

According to the present invention, a variety of routes are contemplated for administration of the methane-oxidizing agent and the pharmaceutical composition. Typically, administration will be accomplished parenterally, but oral administration is also contemplated. Methods of parenteral delivery include topical, intra-arterial, intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, intrauterine, intravaginal, sublingual, or intranasal administration. Preferably, administration is accomplished intraperitoneally and intravenously.

The invention also relates to the in vitro purification of Fe ³⁺ Reduction to Fe ²⁺ Comprising reacting a methane-oxidizing agent as defined above with Fe in a solution ³⁺ The ions are optionally contacted in the presence of a suitable reducing agent.

Preferably, the methane-oxidizing element is a catalyst and a reducing agent is present.

The reducing agent may be nicotinamide adenine Nucleotide (NADH) or water.

The solvent may be a polar protic or aprotic solvent.

Preferably, the solvent and/or reducing agent is water.

Preferably, the reduction reduces 0.5 to 5, more preferably 0.7 to 3, most preferably 1 Fe per methane-oxidizing agent per minute ³⁺ Is performed at a rate of (2).

Preferably, molecular oxygen is produced in the process.

Examples

Example 1: binding ferric iron and reduction to ferrous iron by MB-SB2 (FIG. 1)

50nmol ml of separated out has been measured ^-1 SB2-MB and subsequent addition of 5nmol FeCl ₃ Ultraviolet-visible absorption spectrum of (c).

Example 2: iron reduction (FIG. 2)

Iron reductase activity (1, 2) was determined by a phenanthrazine (Ferrozine) assay.

1.Carter P.1971.Spectrometric determination of serum iron at the submicrogram level with a new reagent(ferrozine).Anal Biochem 40:450-458.

2.Moody MD,Dailey HA.1983.Aerobic ferrisiderophore reductase assay and activity stain for native polyacrylamide gels.Anal Biochem 134:235-239.

Example 3: liver perfusion experiment (figure 3A)

From LPP Atp7b ^/ Rat intubated liver was perfused with Krebs-Ringer bicarbonate solution at 37℃for one hour with 95% O ₂ And 5% CO ₂ And MB-SB2 or MB-OB3b (35. Mu. Mol). During perfusion, total bile was collected at 10 minute intervals. The concentration of iron in bile was determined by inductively coupled plasma emission spectrometry (ICP-OES) as described in (Lichtmannegger et al.j CIin Invest,2016,126,2721-2735).

Example 4: fecal iron excretion (figure 3B)

LPP Atp7b ^-/- Rats were injected intraperitoneally (i.p.) either 110mg/kg body weight MB-SB2 or 150mg/kg body weight MB-OB3b twice daily for four consecutive days. Rats were housed individually in metabolic cages for 4 days. Feces were collected from each rat 24, 48, 72 and 96 hours after the start of treatment over a 24 hour period. Separating feces from food residues, drying, homogenizing by grinding or milling, and concentrating with concentrated HNO ₃ Digestion, iron content was determined by inductively coupled plasma emission spectrometry (ICP-OES).

Example 5: oxidation of Water (FIG. 4)

In a Coy anaerobic chamber (atmosphere 95% Ar 5% H) ₂ ) Preparation of anhydrous FeCl in (Coy Laboratory Products, ann Arbor, MI, USA) ₃ Is a saturated solution of (a). An amount sufficient to produce a 0.5-10 fold excess of metal was added to 100. Mu.l of 1mM-10mM MB-SB2 or MB-OB3b and a headspace gas sample was collected from the vial (head space gas samples). All solutions were 97% H in 0-.8ml brown sealed vials (DWK Life Sciences, milville, N.J., USA) ₂ ¹⁸ O (Sigma Aldrich, st. Louis, mo, USA).

By monitoring ^18,18 O ₂ And H ⁺ Determination of the yield of 2H in the reaction mixture containing metal and MB-SB2 ₂ O is O to O ₂ +4H ⁺ And (5) oxidizing.In the oxygen evolution experiment, at 97% H ₂ ¹⁸ A stock solution of freeze-dried MB-SB2, MB-OB3b, catalase, and anhydrous metal was prepared in O. The reaction mixture contained 2mM MB-SB2 or MB-OB3b and 0.5-20mM metal, with a final volume of 100. Mu.l H ₂ ¹⁸ O. The reaction mixture was prepared in a 2ml brown serum vial and sealed with a polytetrafluoroethylene (Teflon) lined silicon septum. The initial experiment was determined with aluminum foil wrapped vials, but the practice was terminated once it was clear that the same results were produced regardless of whether the vials were wrapped or not. Monitoring of slave H by headspace direct injection (1. Mu.l or 2. Mu.l) ₂ ¹⁸ O production ^18,18 O ₂ 。

Gas samples were manually injected into an Agilent 7890B GC system (Santa Clara, calif., USA) equipped with 7250 Accuate-Mass Q-TOF GC/MS and DB5-MS columns. Except for ^18,18 O ₂ All injections except standard curve injections were performed using a gas tight Hamilton syringe at 1 μl. 97% in 1. Mu.L, 1.5. Mu.L and 2. Mu.L ^18,18 O ₂ (Sigma Aldrich, st Louis, MO, USA) injection produced a standard curve. The headspace in the vial was sampled before and after the addition of the metal, as was the outside air in the mass spectrum as a control. After injection of the standard and control, the samples were mixed and the headspace samples were collected immediately, followed by samples taken every 30-60 seconds. After a few minutes, samples were collected slowly to 1 sample every 2-3 minutes. Generated by ^18,18 O ₂ Is derived from an extracted ion chromatogram set at 35.9978 Da. Observed on some dates ^18,18 O ₂ Is a small offset of the MS position. If it is observed that ^18,18 O ₂ With 97% of MS drift ^18,18 O ₂ The identity of the peak is verified by the standard.

Claims (14)

1. Fe for medicine ³⁺ Reduction of ions to Fe ²⁺ Ionic methane-oxidizing bacteria.

2. The methane-oxidizing rhzomorph for use according to claim 1, wherein the primary structure of the methane-oxidizing rhzomorph comprises a structure according to formula (I):

3. methane-oxidizing mycomycin for use according to claim 1 or 2, wherein Fe is ³⁺ Reduction to Fe ²⁺ An intermediate structure comprising a methane-oxidizing rhzomorph-Fe (II) complex according to formula (II):

4. a methane-oxidizing bacteria element for use according to claims 1 to 3, wherein said methane-oxidizing bacteria element is derived from a methylsporangium strain SB2 (methylbacteria sp.strain SB 2).

5. The methane-oxidizing element for use according to claims 1 to 4, wherein the methane-oxidizing element catalyzes Fe ³⁺ Reduction to Fe ²⁺ And wherein

a) Using water as reducing agent and producing molecular oxygen and/or

b) Reduction is carried out at 0.5 to 5, preferably 0.7 to 3, more preferably 1 Fe per minute per methane-oxidizing mycotin ³⁺ Is performed at a rate of (2).

6. A pharmaceutical composition for use in medicine comprising the methane-oxidizing rhzomorph of claims 1 to 5.

7. The methane-oxidizing bacteria element for use according to claims 1 to 5 or the pharmaceutical composition for use according to claim 6 for use in the treatment of iron overload disorders, neurodegenerative diseases, iron overload disorders caused by red blood cell transfusion, aging, ageing, iron-philic cell death, alcoholic liver disease or amyotrophic lateral sclerosis.

8. The methanotrophic fungus for use according to claim 7 or the pharmaceutical composition for use according to claim 7 for use in the treatment of iron overload disorders.

9. The methane-oxidizing bacteria for use according to claim 7 or 8 or the pharmaceutical composition for use according to claim 7 or 8, wherein the iron overload disorder is caused by a disease selected from the group consisting of hereditary hemochromatosis, thalassemia, sickle cell disease, aplastic anemia, myelodysplasia.

10. The methane-oxidizing element for use according to claim 7 or the pharmaceutical composition according to claim 7, wherein the neurodegenerative disease is selected from alzheimer's disease, parkinson's disease, dementia, huntington's disease, multiple sclerosis.

11. Be used for external Fe ³⁺ Reduction to Fe ²⁺ Comprising reacting a methane-oxidizing rhzomorph as defined in claims 1 to 5 with Fe in solution, optionally in the presence of a suitable reducing agent ³⁺ And (3) ion contact.

12. The method of claim 11, wherein the methane-oxidizing element is a catalyst and a reducing agent is present.

13. The method according to claim 11 or 12, wherein

a) The solvent and/or reducing agent is water and/or

b) Reduction is carried out at 0.5 to 5, preferably 0.7 to 3, more preferably 1 Fe per minute per methane-oxidizing mycotin ³⁺ Is performed at a rate of (2).

14. The method of claim 13, wherein molecular oxygen is produced.