COMPOSITIONS FOR MODULATING GUT MICROBIOTA

Abstract

The present invention provides compositions that modulate gut microbiota and/or the activity of deoxyribonucleases (e.g., in the gastrointestinal tract) and are useful in the treatment of diseases and disorders related to the activity of deoxyribonucleases including, for example, neurodegenerative, inflammatory, and metabolic diseases and cancer.

Claims

1. A product for maintenance of DNase activity, the product comprising Mg.sup.2+ and Na.sup.+ cations and water, wherein the product is substantially free of cations other than Mg.sup.2+ and Na.sup.+ cations and/or other cations are in mass with other chemical components of water by mass less than 50 mg/l.

2. The product of claim 1, further comprising an additive selected from the group consisting of: ascorbic acid; B vitamins; extracts or infusions of green tea; aloe; garcinia; gentiana; corn silk; turmeric; grapefruit; and citric acid.

3. The product of claim 1, wherein the Na.sup.+ is derived from sodium bicarbonate or sodium sulfate, and wherein the Mg.sup.2+ is derived from magnesium chloride or magnesium oxide and/or succinic acid is added.

4. (canceled)

5. The product of claim 1, wherein the product is essentially free of any cation other than sodium and magnesium.

6. The product of claim 1, comprising about 100.0 mg/l Mg.sup.2+ to about 140.0 mg/l Mg.sup.2+.

7. (canceled)

8. (canceled)

9. (canceled)

10. The product of claim 1, comprising about 0.1 mg/l Na.sup.+ to about 2.0 mg/l Na.sup.+.

11. The product of claim 1, comprising about 6.0 to 15.0 mg/l Mg.sup.2+.

12. (canceled)

13. (canceled)

14. (canceled)

15. (canceled)

16. The product of claim 1, wherein said composition is at room temperature.

17. (canceled)

18. The product of claim 1, wherein the water is selected from demineralized water or distilled water.

19. (canceled)

20. A method of modulating activity of a DNase comprising contacting the DNase with the product of claim 1.

21. A method of modulating activity of a DNase in a subject, comprising administering to the subject the product of claim 1.

22. The method of claim 21, wherein the modulating increases DNase activity in the gastrointestinal tract of the subject, and wherein the DNase is deoxyribonuclease I or deoxyribonuclease II.

23. (canceled)

24. (canceled)

25. A method of treating a disease or disorder associated with DNase activity in a subject in need thereof comprising administering to the subject the product of claim 1.

26. The method of claim 25, wherein the disease or disorder is selected from the group consisting of neurodegenerative disorders; diabetes; cancer; graft versus host disease; systemic lupus erythematosus; metabolic syndrome; weight gain and obesity.

27. The method of claim 26, wherein the diabetes is type 1 diabetes.

28. The method of claim 26, wherein the cancer treatment involves maintaining the condition of patients undergoing chemotherapy or X-ray therapy.

29. (canceled)

30. The method of claim 26, wherein the neurodegenerative disease or disorder is selected from the group consisting of Alzheimer's disease; Parkinson's disease; dementia and CADASIL syndrome.

31. The method of claim 25, wherein the disease or disorder is selected from the group consisting of an enteric viral disease, a bacteriophage disorder, and a metabolic syndrome.

32. The method of claim 25, wherein the administration is conducted three times a day for at least one week.

33. The method of claim 25, where approximately 200 mL of the composition is administered in each administration.

34. (canceled)

35. (canceled)

Description

DETAILED DESCRIPTION

[0016] This invention provides water compositions that are highly effective for modulating the activity of the deoxyribonuclease (DNase) family of enzymes. In particular, this invention provides products for maintenance of DNase activity, that contain Mg.sup.2+ and Na.sup.+ cations and water, while other cations are either absent or in mass with other chemical components of water by mass less than 50 mg/l.

[0017] The Mg.sup.2+ can be derived from any suitable magnesium source known to those of ordinary skill, such as magnesium chloride or magnesium oxide. Further, citric acid and/or succinic acid can be used to increase the solubility of magnesium oxide. Suitable concentrations of magnesium cation include, but are not limited to, e.g., about 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 20.0, 30.0, 40.0, 50.0, 60.0, 70.0, 80.0, 90.0, 100.0, 110.0, 120.0, 130.0, 140.0, 150.0 or about 155.0 mg/l Mg.sup.2+, or about 6.0 to about 15.0 mg/l Mg.sup.2+, or about 140.0 mg/l Mg.sup.2+.

[0018] The Na.sup.+ can be derived from any suitable sodium source known to those of ordinary skill in the art, such as sodium bicarbonate or sodium sulfate. Suitable concentrations of sodium include, but are not limited to, e.g., about 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, or about 12 mg/l Na.sup.+, or about 0.1 or about 0.5 or about 2.0 mg/l Na.sup.+.

[0019] In some embodiments, the water composition comprises: about 6.0 to 15.0 mg/l Mg.sup.2+, about 12 mg/l Na.sup.+; and water.

[0020] In some embodiments of the invention, the composition is essentially free of any cation other than sodium and magnesium cations. In some embodiments, the composition comprises only sodium and magnesium cations in water and no other components.

[0021] In some embodiments of the invention, the water composition consisting essentially of: (i) about 6.0 to 155.0 mg/l Mg.sup.2+; (ii) about 0.005 to 12 mg/l Na.sup.+; and (iii) water, such that there are no additional components added to the composition which materially affect the basic and novel characteristics of the invention.

[0022] In some embodiments of the invention, the water composition consists of (i) about 6.0 to 155.0 mg/l Mg.sup.2+; (ii) about 0.005 to 12 mg/l Na.sup.+; and water, such that no other components are added to the composition.

[0023] In some embodiments, the water composition is provided at room temperature. The water compositions may further comprise an additive selected from ascorbic acid; B vitamins; extracts or infusions of green tea; aloe; garcinia; gentiana; corn silk; turmeric; grapefruit; and citric acid.

[0024] The water used in the compositions may include certain forms of water such as very low mineral content water (or light mineral water) having a mineral concentration of less than 50 mg/l. Demineralized water and distilled water may also be used.

[0025] Without being bound by any particular theory of the invention, Applicants believe that the composition of the invention modulates DNase activity and, in turn: (i) prevents the formation of DNA protein complexes in the microbiota, (ii) promotes the maintenance of barrier functions of the gut (iii) reduces the penetration of bacteria, bacteriophages, microbial toxins and matrix components into the systemic bloodstream, (iv) inhibits the development of neurodegenerative and oncological diseases, diabetes, metabolic syndrome, (v) helps to reduce excess body weight, and (vi) improves the condition of patients subjected to with chemotherapy and X-ray therapy.

Methods

[0026] Compositions of the invention can modulate activity of one or more deoxyribonucleases (DNases). The term “modulate” is meant to refer to an ability to increase or decrease the activity of one or more members of the DNase family of enzymes. Accordingly, compositions of the invention can be used in methods of modulating a DNase by contacting the DNase with any one or more of the compositions described herein. In some embodiments, compositions of the present invention can act as inhibitors of one or more DNases. In some embodiments, compositions of the present invention can act to stimulate the activity of one or more DNases. In further embodiments, the compositions of the invention can be used to modulate activity of a DNase in an individual in need of modulation of the enzyme by administering a modulating amount of a composition of the invention.

[0027] DNases to which the present compositions interact with and/or modulate include any member of the DNase family, in particular the Mg-dependent DNase family. In some embodiments, the DNase is deoxyribonuclease I.

[0028] Another aspect of the present invention pertains to methods of treating a DNase-associated disease or disorder in an individual (e.g., patient) by administering to the individual in need of such treatment a therapeutically effective amount or dose of a composition of the present invention. A DNase-associated disease can include any disease, disorder or condition that is directly or indirectly linked to expression or activity of the DNase, including overexpression and/or abnormal activity levels. A DNase-associated disease can also include any disease, disorder or condition that can be prevented, ameliorated, or cured by modulating DNase activity.

Examples of DNase-associated diseases include diseases involving the immune system including, for example, organ transplant rejection (e.g., allograft rejection and graft versus host disease). Further examples of DNase-associated diseases include autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, juvenile arthritis, diabetes (e.g., type I diabetes), lupus, psoriasis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, or autoimmune thyroid disorders. Further examples of DNase-associated diseases include allergic conditions such as asthma, food allergies, atopic dermatitis and rhinitis. Further examples of DNase-associated diseases include viral diseases such as Epstein Barr Virus (EBV).sub.5 Hepatitis B, Hepatitis C.sub.5 HIV.sub.5 HTLV I.sub.5 Varicella-Zoster Virus (VZV) and Human Papilloma Virus (HPV). Further examples of DNase-associated diseases or conditions include skin disorders such as atopic dermatitis, psoriasis (for example, psoriasis vulgaris), skin sensitization, and the like. In further embodiments, the DNase-associated disease is cancer such as, for example, prostate, renal, hepatocellular, pancreatic, gastric, breast, lung, cancers of the head and neck, glioblastoma, leukemia, lymphoma or multiple myeloma.

[0029] In some embodiments, the disease or disorder is a neurodegenerative disorder, such as Alzheimer's disease, Parkinson's disease, dementia, or CADASIL syndrome. In some embodiments, the disease or disorder is diabetes, such as type 1 diabetes. In some embodiments, the disease or disorder is cancer. The cancer treatment may involve maintaining the condition of patients undergoing chemotherapy or X-ray therapy.

[0030] In some embodiments, the disease or disorder is graft versus host disease.

[0031] In some embodiments, the disease or disorder is systemic lupus erythematosus.

[0032] In some embodiments, the disease or disorder is metabolic syndrome.

[0033] In some embodiments the disease or disorder is weight gain.

[0034] In some embodiments, the disease or disorder is obesity.

[0035] In some embodiments, the disease or disorder is an enteric viral disease.

[0036] In some embodiments, the disease or disorder is a bacteriophage disorder.

[0037] In some embodiments, the disease or disorder is a metabolic syndrome.

[0038] In some embodiments, the enteric viral disease involves, for example, an infection caused by Hepatitis A virus, Norovirus, Rotavirus, Hepatitis E virus, Astrovirus, Reovirus, or Echovirus.

[0039] In some embodiments, the metabolic syndrome includes, for example, conditions such as obesity, fatty liver, diabetes, hyperlipidemia, hypertension, hypercholesterolemia, hyper low-density lipoprotein (LDL) cholesterinosis, cardiovascular disease, arteriosclerosis, and coronary artery disease.

[0040] As used herein, the term “contacting” refers to the bringing together of indicated moieties in an in vitro system or an in vivo system. For example, “contacting” a DNase with a composition of the invention includes the administration of a composition of the present invention to an individual or patient, such as a human, having a DNase, as well as, for example, introducing a composition of the invention into a sample containing a cellular or purified preparation containing the DNase.

[0041] As used herein, the term “subject” or “individual” or “patient,” used interchangeably, refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans.

[0042] As used herein, the phrase “therapeutically effective amount” refers to the amount of active composition or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following:

[0043] (1) preventing the disease; for example, preventing a disease, condition or disorder in an individual who may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease; (2) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology), and (3) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology).

[0044] One or more additional pharmaceutical agents such as, for example, chemotherapeutics, anti-inflammatory agents, and/or immunosuppressants can be used in combination with the compositions of the present invention for treatment of DNase-associated diseases, disorders or conditions. For example, the inventive composition may be used in combination with a chemotherapeutic in the treatment of colorectal cancer (or some other form of cancer) may improve the treatment response as compared to the response to the chemotherapeutic agent alone, without exacerbation of its toxic effects. Additive or synergistic effects are desirable outcomes of combining a composition of the present invention with an additional agent. The agents can be combined with the present compositions in a single or continuous dosage form, or the agents can be administered simultaneously or sequentially as separate dosage forms.

Kits

[0045] The present invention also includes pharmaceutical kits useful, for example, in the treatment or prevention of DNase-associated diseases or disorders, such as cancer, which include one or more containers containing a solution comprising a therapeutically effective amount of a composition described herein. Such kits can further include, if desired, one or more of various conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to those skilled in the art. Instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and/or guidelines for mixing the components, can also be included in the kit.

[0046] The invention can be described and demonstrated by way of specific examples. The following examples are offered for illustrative purposes and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same results.

EXAMPLES

[0047] Reagents and solvents are obtained from commercial sources such as Sigma-Aldrich.

Example 1

[0048] This example describes the preparation of a product of the present invention that is useful for the maintenance of DNase activity. To a sample of very low mineral content water (light mineral water; demineralized water or distilled water may also be used) was added Mg.sup.2+ (in the form of magnesium chloride or magnesium oxide) and Na.sup.+ (in the form of sodium bicarbonate or sodium sulfate) to a final concentration of 7.0-155.0 mg/l, and 0.005-12.0 mg/l, respectively. The pH of the water composition was adjusted to a final pH of 6.0 to 7.5.

TABLE-US-00001 Reagent Amount Mg.sup.2+ (MgCl.sub.2)  7.0-155.0 mg/l Na.sup.+ (NaHCO.sub.3) 0.005-12.0 mg/l Water Balance

Example 2

[0049] This example describes a method for determining DNase activity in Alzheimer's disease patients.

[0050] In this experiment, five patients (male), aged 72±2 years, with Alzheimer's disease were studied. To each patient was administered a water composition of the invention having the following constitution:

TABLE-US-00002 Reagent Amount Mg.sup.2+ (MgO) 100.0 mg/l Na.sup.+ (Na.sub.2SO.sub.4)  2.00 mg/l Demineralized water Balance
The composition (200 mL) was administered to each patient three times a day for one week. During this experiment, fecal samples were collected from each patient using a sterile collection tube. The samples (10 mg feces) were diluted in phosphate buffered saline (PBS) (0.02 mol/L, pH 7.2-7.4) (100 μl PBS). Samples were fully shaken and then left standing for 10 minutes. The samples were then centrifuged for approximately 20 minutes at 3000 rpm. The supernatants from each sample were collected and either assayed immediately or stored at −80° C. DNase activity was determined using standard colorimetric methods (Sinicropi, D., Baker, D. L., Prince, W. S., Shiffer, K., Shak, S., Analytical Biochemistry, “Colorimetric Determination of DNase I Activity with a DNA Methyl Green Substrate,” Volume 222, Issue 2, 1 Nov. 1994, Pages 351-358). The data from this experiment is shown in Table 1.

TABLE-US-00003 TABLE 1 DNase Activity in Alzheimer's Disease Patients. DNase activity Patient Before One week later 1  ND* 0.15 2 ND 0.20 3 0.100 0.20 4 ND 0.1 5 ND 0.25 *ND = not detected

Example 3

[0051] This example describes a method for determining DNase activity in patients that have colorectal cancer. In this experiment, three patients (male), aged 57±6 years, with colorectal cancer were studied. To each patient was administered a water composition of the invention having the following constitution:

TABLE-US-00004 Reagent Amount Mg.sup.2+ (MgCl.sub.2) 140.0 mg/l Na.sup.+ (NaHCO.sub.3)  0.10 mg/l Distilled water Balance

[0052] The composition (200 mL) was administered to each patient three times a day for one week. Fecal samples were collected as described in Example 2. The data from this experiment is shown in Table 2.

TABLE-US-00005 TABLE 2 DNase Activity in Colorectal Cancer Patients. DNase activity Patient Before One week later 1  ND* 0.125 2 ND 0.150 3 ND 0.120 *ND = not detected

Example 4

[0053] This example describes a method for determining DNase activity in patients that have metabolic syndrome. Two groups of three patients (male), aged 68±5 years, with metabolic syndrome were studied. The patients in the study were overweight by 29.0±6.0 kg. To each patient was administered a water composition of the invention having the following constitution:

TABLE-US-00006 Reagent Amount (Group 1) Amount (Group 2) Mg.sup.2+ (MgCl.sub.2) 140.0 mg/l 8.0 mg/l Na.sup.+ (NaHCO.sub.3)  0.50 mg/l 0.5 mg/l Distilled water Balance Balance
The composition (200 mL) was administered to each patient three times a day for a period of four weeks. Fecal samples were collected as described in Example 2. The data from this experiment is shown in Table 3.

TABLE-US-00007 TABLE 3 DNase Activity in Patients with Metabolic Syndrome. Uric acid level DNase activity Weight (Kg) (mg/dL) 4 weeks 4 weeks 4 weeks Patient Before later Before later Before later Group 1 1 0120 0.250 82.0 79.5 8.2 7.5 2 0.115 0.190 97.0 94.0 8.62 7.85 3 0.220 0.280 102.0 99.0 7.85 6.10 Group 2 4 0.110 0.150 98.0 95.0 9.0 8.2 5 0.200 0.240 114.0 110.0 8.7 6.93 6 ND* 0.200 96.0 93.5 7.9 6.3 *ND = not detected

[0054] These data indicate that in patients with metabolic syndrome, the composition of the invention is capable of causing an increase in DNase activity in feces, a decrease level of serum uric acid and a decrease in body weight after an administration period of one month.