LACTOBACILLUS PLANTARUM COMPOSITIONS AND USES THEREOF

Abstract

The invention relates to at least one probiotic strain of Lactobacillus plantarum for use in the treatment and/or prevention of age-related systemic inflammation in a human.

Claims

1. A method for treating and/or preventing age-related systemic inflammation in a human, comprising administering to a human in need thereof a therapeutically effective dose of at least one probiotic strain of Lactobacillus plantarum.

2. The method according to claim 1, wherein the human is aged more than 60, 65, 70, 75, 80, 85 or 90 years.

3. The method according to claim 1 or 2, wherein the human is a man.

4. The method according to claim 1 or 2, wherein the human is a woman.

5. The method according to claim 4, wherein the woman is a post-menopausal woman.

6. The method according to any one of claims 1-5, wherein the effective dose of at least one probiotic strain of Lactobacillus plantarum is administered at least once a day.

7. The method according to any one of claims 1-6, wherein the effective dose of the at least one probiotic strain of Lactobacillus plantarum is from about 10.sup.6 to about 10.sup.14 colony forming units (CFU) per dose, preferably from about 10.sup.8 to about 10.sup.12 CFU per dose, or more preferably from about 10.sup.9 to about 10.sup.11 CFU per dose.

8. The method according to any one of claims 1-7, wherein one or more effective doses of the at least one probiotic strain of Lactobacillus plantarum are administered in one day, and wherein the daily dose of the at least one probiotic strain of Lactobacillus plantarum is from about 10.sup.6 to about 10.sup.14 CFU per day, preferably from about 10.sup.8 to about 10.sup.12 CFU per day, or more preferably from about 10.sup.9 to about 10.sup.11 CFU per day.

9. The method according to any one of claims 1-8, wherein the treatment and/or prevention of age-related systemic inflammation involves reducing and/or preventing an increase in the level of C-reactive protein (CRP) and/or reducing and/or preventing an increase in the level of calprotectin.

10. The method according to any one of claims 1-9, wherein the at least one probiotic strain of Lactobacillus plantarum is chosen from Lactobacillus plantarum 299 (DSM 6595), Lactobacillus plantarum 299v (DSM 9843), Lactobacillus plantarum HEAL 9 (DSM 15312), Lactobacillus plantarum HEAL 19 (DSM 15313), Lactobacillus plantarum HEAL 99 (DSM 15316) and Lactobacillus plantarum GOS42 (DSM 32131).

11. The method according to claim 10, wherein the at least one probiotic strain of Lactobacillus plantarum is Lactobacillus plantarum HEAL 9 (DSM 15312).

12. The method according to any one of claims 1-11, wherein the at least one probiotic strain is administered in a composition comprising at least one carrier selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable excipient, a diluent, and a food.

13. The method according to claim 12, wherein the composition is provided in the form of a solution, suspension, emulsion, tablet, granule, powder, capsule, lozenge, chewing gum, or suppository.

14. The method according to claim 12, wherein the food is a cereal-based product, a dairy product, a juice drink, or a fermented food.

15. At least one probiotic strain of Lactobacillus plantarum for use in the treatment and/or prevention of age-related systemic inflammation in a human.

16. A pharmaceutical composition comprising the at least one probiotic strain according to claim 15, and one or more pharmaceutically acceptable excipients, for use in the treatment and/or prevention of age-related systemic inflammation in a human.

17. Use of a composition comprising at least one probiotic strain according to claim 15, or use of a pharmaceutical composition according to claim 16, in the treatment and/or prevention of age-related systemic inflammation in a human.

18. A method or use for the treatment of age-related systemic inflammation in a human as claimed in any one of claims 1 to 17 wherein the human has systemic inflammation indicated by a serum CRP level of from 2-10 mg/L or 3-10 m/L.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0080] FIG. 1 shows the mean change in actual/absolute mg/L serum values of C-reactive protein (CRP) levels from baseline, after four weeks of treatment, for each of the three treatment groups.

EXEMPLARY DOSAGE FORMS

[0081] In addition to the formulations referenced above (and incorporated herein by reference), the following examples illustrate pharmaceutical formulations according to the invention.

Example A: Tablet

[0082]

TABLE-US-00001 Probiotic strain(s) 1 × 10.sup.9 CFU Lactose 200 mg Starch 50 mg Polyvinylpyrrolidone 5 mg Magnesium stearate 4 mg

[0083] Tablets are prepared from the foregoing ingredients by wet granulation followed by compression.

Example B: Tablet Formulations

[0084] The following formulations A and B are prepared by wet granulation of the ingredients with a solution of povidone, followed by addition of magnesium stearate and compression.

Formulation A

[0085]

TABLE-US-00002 (a) Probiotic strain(s) 1 × 10.sup.9 CFU 1 × 10.sup.9 CFU (b) Lactose B.P. 210 mg 26 mg (c) Povidone B.P. 15 mg 9 mg (d) Sodium Starch Glycolate 20 mg 12 mg (e) Magnesium Stearate 5 mg 3 mg

Formulation B

[0086]

TABLE-US-00003 (a) Probiotic strain(s) 1 × 10.sup.9 CFU 1 × 10.sup.9 CFU (b) Lactose 150 mg — (c) Avicel PH 101 ® 60 mg 26 mg (d) Povidone B.P. 15 mg 9 mg (e) Sodium Starch Glycolate 20 mg 12 mg (f) Magnesium Stearate 5 mg 3 mg

Formulation C

[0087]

TABLE-US-00004 Probiotic strain(s) 1 × 10.sup.9 CFU Lactose 200 mg Starch 50 mg Povidone 5 mg Magnesium stearate 4 mg

[0088] The following formulations, D and E, are prepared by direct compression of the admixed ingredients. The lactose used in formulation E is of the direction compression type.

Formulation D

[0089]

TABLE-US-00005 Probiotic strain(s) 1 × 10.sup.9 CFU Pregelatinised Starch NF15 150 mg

Formulation E

[0090]

TABLE-US-00006 Probiotic strain(s) 1 × 10.sup.9 CFU Lactose 150 mg Avicel  ® 100 mg

Formulation F (Controlled Release Formulation)

[0091] The formulation is prepared by wet granulation of the ingredients (below) with a solution of povidone followed by the addition of magnesium stearate and compression.

TABLE-US-00007 (a) Probiotic strain(s) 1 × 10.sup.9 CFU (b) Hydroxypropylmethylcellulose 112 mg (Methocel K4M Premium) ® (c) Lactose B.P. 53 mg (d) Povidone B.P.C. 28 mg (e) Magnesium Stearate 7 mg

[0092] Release takes place over a period of about 6-8 hours and was complete after 12 hours.

Example C: Capsule Formulations

Formulation A

[0093] A capsule formulation is prepared by admixing the ingredients of Formulation D in Example B above and filling into a two-part hard gelatin capsule. Formulation B (infra) is prepared in a similar manner.

Formulation B

[0094]

TABLE-US-00008 (a) Probiotic strain(s) 1 × 10.sup.9 CFU (b) Lactose B.P. 143 mg (c) Sodium Starch Glycolate 25 mg (d) Magnesium Stearate 2 mg

Formulation C

[0095]

TABLE-US-00009 (a) Probiotic strain(s) 1 × 10.sup.9 CFU (b) Macrogol 4000 BP 350 mg

[0096] Capsules are prepared by melting the Macrogol 4000 BP, dispersing the probiotic strain(s) in the melt and filling the melt into a two-part hard gelatin capsule.

Formulation D (Controlled Release Capsule)

[0097] The following controlled release capsule formulation is prepared by extruding ingredients a, b, and c using an extruder, followed by spheronisation of the extrudate and drying. The dried pellets are then coated with release-controlling membrane (d) and filled into a two-piece, hard gelatin capsule.

TABLE-US-00010 (a) Probiotic strain(s) 1 × 10.sup.9 CFU (b) Microcrystalline Cellulose 125 mg (c) Lactose BP 125 mg (d) Ethyl Cellulose 13 mg

Experimental Example 1

Materials and Methods

[0098] The possible anti-inflammatory activity of the probiotic product was evaluated in a randomized double-blind placebo-controlled trial with 66 healthy participants>70 years of age with low grade systemic inflammation (defined by C-reactive protein; serum level 2-10 mg/L).

[0099] Criteria for exclusion from the study were: [0100] Intake of antibiotic treatment in the last four weeks before inclusion into the study; [0101] Currently on corticosteroid treatment; [0102] Presence of chronic inflammatory disease.

[0103] The subjects were randomly allocated to one of the three groups: [0104] 1. Lactobacillus plantarum Heal 9 (Lp Heal 9) [0105] 2. Lactobacillus plantarum Heal 9+berries (Bär+Lp Heal 9) [0106] 3. Placebo

[0107] Each study product was formulated as a powder at 10 g/dose and was to be mixed with sour milk/yoghurt and consumed once daily for a period of four weeks. Test Product A (for group 2) consisted of a daily dose of 1 billion colony forming units (10.sup.9 CFU/dose) of freeze-dried Lactobacillus plantarum HEAL 9 probiotic bacteria, freeze dried berries (blackberries and blackcurrants) and maltodextrin. Test Product B (for group 1) consisted of a daily dose of 1 billion colony forming units (10.sup.9 CFU/dose) of freeze-dried Lactobacillus plantarum HEAL 9 probiotic bacteria, and maltodextrin, treated to resemble Test Product A in appearance and taste. The placebo product consisted of maltodextrin, treated with colourants and flavourings/aromatic agents to resemble the Test Product A in appearance and taste.

[0108] The participants were also asked to keep a study diary throughout the study period for the documentation of their intestinal health and as a means for checking compliance and to refrain from taking other products containing probiotic bacteria.

[0109] Blood and faecal samples were taken at baseline and at the end of the study for the analysis of the following parameters: [0110] 1. Faecal samples were used for the analysis of calprotectin (a marker of gut inflammation) and zonulin (a protein that modulates the permeability of tight junctions between cells of the intestinal wall and is used as a marker of increased gut permeability); [0111] 2. Blood samples were used for the analysis of the systemic inflammation markers CRP and fibrinogen.

[0112] CRP levels in blood, serum and plasma may be determined using commercially available methods and apparatus, such as the Alere Afinion™ CRP assay using the Afinion™ AS100 analyser from Alere/Abbott (see www.alere.com).

[0113] This test is an in vitro method using a solid phase immunochemical assay based on a membrane coated with anti-human CRP antibodies, which react with CRP in the sample.

[0114] The analyser measures the colour intensity of the membrane, and this is proportional to the amount of CRP in the sample.

[0115] CRP levels in serum can be tested in a sensitive manner by a variety of methods (see Pearson T A et al (2003) Markers of inflammation and cardiovascular disease: application to clinical and public health practice: A statement for healthcare professionals from the Centers for Disease Control and Prevention and the American Heart Association. Circulation. 107; 499-511

Results

[0116] The Wilcoxon Rank-sum Test was used for statistical analysis in the study.

[0117] No differences in the levels of zonulin and fibrinogen were detected between the probiotic groups and the placebo.

[0118] However, the level of the inflammatory marker CRP increased over time in the placebo group (group 3) and reduced in the group receiving Lp HEAL 9 and berries—Bär+Lp Heal 9 (group 2) (FIG. 1). The effect was even more pronounced in the group consuming only probiotics, without the addition of berries (group 1) (FIG. 1).

[0119] The level of calprotectin expressed as mean change over time did not differ between either of the probiotic groups and placebo. However, there were significantly fewer participants in the Lactobacillus plantarum (Lp HEAL 9 only group (group 1) that showed increased levels for calprotectin over time compared to placebo (group 3) (p=0.028) (Table 1).

TABLE-US-00011 TABLE 1 Analysis of the number of participants with stable or reduced levels of calprotectin vs increased levels of calprotectin Participants with stable Participants with or reduced levels of increased levels of p- calprotectin (% of group) calprotectin (% of group) value Group 1: 15 (83.3) 3 (16.6) 0.028 Lp HEAL 9 Group 3: 11 (50)   11 (50)   Placebo

CONCLUSION

[0120] The results obtained with CRP and calprotectin show that Lactobacillus plantarum, in particular Lactobacillus plantarum HEAL 9, has efficacy in treating and/or preventing age-related systemic inflammation in otherwise healthy elderly people.