LACTOBACILLUS COMPOSITIONS AND USES THEREOF

Abstract

The invention relates to the use of at least one strain of Lactobacillus plantarum to increase vigour in a human.

Claims

1. At least one strain of Lactobacillus plantarum for use in a method of increasing vigour in a human compared to their vigour without treatment, comprising treatment by administering to a human at least one strain of Lactobacillus plantarum.

2. Use of at least one strain of Lactobacillus plantarum to increase vigour in a human.

3. A method of increasing vigour in a human compared to their vigour without treatment, the method comprising treatment by administering to a human at least one strain of Lactobacillus plantarum.

4. A use, at least one strain for use, or a method as claimed in any of claims 1 to 3, wherein the human to be treated is in need of increased vigour.

5. A use, at least one strain for use, or a method as claimed in claim 4, wherein the human to be treated has iron deficiency.

6. A use, at least one strain for use, or a method as claimed in any of claims 1 to 5, wherein the human is an athlete.

7. A use, at least one strain for use, or a method as claimed in any of claims 1 to 6, wherein the human is female, preferably a female athlete.

8. A use, at least one strain for use, or a method as claimed in claim 7, wherein the female is premenopausal.

9. A use, at least one strain for use, or a method according to any one of claims 1 to 8, wherein the at least one strain is administered at a daily dose of from about 1×10.sup.6 to about 1×10.sup.14 colony forming units (CFU).

10. A use, at least one strain for use, or a method according to any one of claims 1 to 9, wherein the daily dose of the at least one strain is from about 1×10.sup.9 to about 1×10.sup.11 CFU.

11. A use, at least one strain for use, or a method according to claim 10, wherein the daily dose of the at least one strain is 1×10.sup.10 CFU.

12. A use, at least one strain for use, or a method according to any one of claims 1 to 11, wherein the at least one strain is administered at a daily dose in a single unit dosage form.

13. A use, at least one strain for use, or a method according to any one of claims 9 to 12, wherein the daily dose of at least one strain is administered for at least four weeks.

14. A use, at least one strain for use, or a method according to claim 13, wherein the daily dose of at least one strain is administered for at least eight weeks.

15. A use, at least one strain for use, or a method according to claim 14, wherein the daily dose of at least one strain is administered for at least twelve weeks.

16. A use, at least one strain for use, or a method according to any preceding claim, wherein the at least one strain of Lactobacillus plantarum is selected from Lactobacillus plantarum DSM 15312, Lactobacillus plantarum DSM 15313, Lactobacillus plantarum DSM 15316, Lactobacillus plantarum DSM 6595, Lactobacillus plantarum DSM 9843, Lactobacillus plantarum DSM 32131, Lactobacillus plantarum DSM 17852, and Lactobacillus plantarum DSM 17853.

17. A use, at least one strain for use, or a method according to any preceding claim, wherein the at least one strain of Lactobacillus plantarum is able to adhere to the intestinal epithelium and persist in the intestine.

18. A use, at least one strain for use, or a method according to any preceding claim, wherein the at least one strain of Lactobacillus plantarum comprises a mannose-specific adhesin.

19. A use, at least one strain for use, or a method according to any preceding claim, wherein the at least one strain of Lactobacillus plantarum is Lactobacillus plantarum DSM 9843 (299v®).

20. A use, at least one strain for use, or a method according to any preceding claim, wherein the at least one strain is administered in the form of a composition comprising at least one pharmaceutically and/or nutritionally acceptable carrier, excipient and/or diluent material.

21. A use, at least one strain for use, or a method according to claim 20, wherein the carrier material is a food.

22. A use, at least one strain for use, or a method according to claim 20 or 21, wherein the composition is provided in a form selected from a solution, suspension, emulsion, tablet, granule, powder, capsule, lozenge, chewing gum, or suppository.

23. A use, at least one strain for use, or a method according to claim 22, wherein the composition is provided in the form of capsule.

24. A use, at least one strain for use, or a method according to any of claims 20 to 23, wherein the composition comprises iron.

25. A use, at least one strain for use, or a method according to claim 24, wherein the iron is provided in the form of an iron salt.

26. A use, at least one strain for use, or a method according to claim 25, wherein the iron salt is ferrous fumarate.

27. A use, at least one strain for use, or a method according to any of claims 24 to 26 wherein the composition is provided in a capsule and contains Lactobacillus plantarum DSM 9843 (299v®) at a concentration of 1×10.sup.10 CFU/capsule and 20 mg iron, preferably ferrous-fumarate.

28. A use, at least one strain for use, or a method according to any preceding claim wherein increased vigour is measured using a Profile of Mood States (POMS) psychological rating scale.

29. A use, at least one strain for use, or a method as claimed in claim 30, wherein the increased vigour by treatment with at least one strain of Lactobacillus plantarum is not accompanied by improvements in one, two, three, four or all five of the other mood states of Tension, Depression, Anger, Fatigue, and Confusion, according to a POMS psychological rating scale.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0068] FIG. 1: Graph plotting POMS test score against mood states, prior to competition, for successful and less successful performers from Morgan & Johnson (1978)-supra.

TABLE-US-00001 TABLE 1 online POMS Questionnaire (Mackenzie B. (2001) supra) Feeling How I have felt** Friendly Tense Angry Worn Out Unhappy Clear Headed Lively Confused Sorry for things done Shaky Listless Peeved Considerate Sad Active On Edge Grouchy Blue Energetic Panicky Hopeless Relaxed Unworthy Spiteful Sympathetic Uneasy Restless Unable to Concentrate Fatigued Helpful Annoyed Discouraged Resentful Nervous Lonely Miserable Muddled Cheerful Bitter Exhausted Anxious Ready to Fight Good Natured Gloomy Desperate Sluggish Rebellious Helpless Weary Bewildered Alert Deceived Furious Efficient Trusting Full of Pep Bad Tempered Worthless Forgetful Carefree Terrified Guilty Vigorous Uncertain about things Bushed **Selection from “Not at All”; “A Little”; “Moderately”; “Quite a Bit”; “Extremely”.

[0069] Assessment

[0070] A Total Mood Disturbance (TMD) score (possible range of −32 to 200) is calculated by adding scores for Tension (0-36), Depression (0-60), Anger (0-48), Fatigue (0-28) and Confusion (0-28) and then subtracting the Vigour score (0-32).

[0071] The scores in brackets (x-y) above indicate the possible score range with lower scores indicative of people with more stable mood profiles.

[0072] Normative Data

[0073] TERRY, P. (n.d.) Normative Values for the Profile of Mood States for Use with Athletic Samples, [WWW] Available from: https://eprints.usq.edu.au/4385/2/Terry_Lane_JASS_v12n1_Authors_version.pdf [Accessed Jun. 18, 2019]) provides POMS norms for an athletic sample (n=2086) grouped by level of competition (International standard athletes, club level athletes and recreational athletes).

TABLE-US-00002 Group Tension Depression Anger Vigour Fatigue Confusion International 5.66 4.38 6.24 18.51 5.37 4.00 Club 9.62 8.67 9.91 15.64 8.16 7.38 Recreational 6.00 3.11 3.60 17.78 6.37 4.84

[0074] Analysis

[0075] Analysis of the POMS result is by comparing it with the results of previous POMS tests. Morgan & Johnson (MORGAN, W. P. and JOHNSON, R. W. (1978) “Personality characteristics of successful and unsuccessful oarsmen.” International Journal of Sport Psychology, 9, p. 119-133) found that plotting the mood state results of elite performers prior to competition exhibited the FIG. 1 graph. This graph, with a raised peak for Vigour, was termed the “Iceberg” profile, identifying successful performers.

[0076] Exemplary Dosage Forms

[0077] In addition to the formulations referenced above (and incorporated herein by reference), the following examples illustrate pharmaceutical formulations and other formulations according to the invention.

[0078] Example A: Tablet

TABLE-US-00003 Lactobacillus strain(s) 1 × 10.sup.10 CFU Lactose 200 mg Starch 50 mg Polyvinylpyrrolidone 5 mg Magnesium stearate 4 mg

[0079] Tablets are prepared from the foregoing ingredients by wet granulation followed by compression.

[0080] Example B: Tablet Formulations

[0081] 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.

[0082] Formulation A

TABLE-US-00004 (a) Lactobacillus strain(s) 1 × 10.sup.10 CFU 1 × 10.sup.10 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

[0083] Formulation B

TABLE-US-00005 (a) Lactobacillus strain(s) 1 × 10.sup.10 CFU 1 × 10.sup.10 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

[0084] Formulation C

TABLE-US-00006 Lactobacillus strain(s) 1 × 10.sup.10 CFU Lactose 200 mg Starch 50 mg Povidone 5 mg Magnesium stearate 4 mg

[0085] 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.

[0086] Formulation D

TABLE-US-00007 Lactobacillus strain(s) 1 × 10.sup.10 CFU Pregelatinised Starch NF15 150 mg

[0087] Formulation E

TABLE-US-00008 Lactobacillus strain(s) 1 × 10.sup.10 CFU Lactose 150 mg Avicel ® 100 mg

[0088] Formulation F (Controlled Release Formulation) 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-00009 (a) Lactobacillus strain(s) 1 × 10.sup.10 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

[0089] Release takes place over a period of about 6-8 hours and was complete after 12 hours.

[0090] Example C: Capsule Formulations

[0091] Formulation A 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.

[0092] Formulation B

TABLE-US-00010 (a) Lactobacillus strain(s) 1 × 10.sup.10 CFU (b) Lactose B.P. 143 mg (c) Sodium Starch Glycolate 25 mg (d) Magnesium Stearate 2 mg

[0093] Formulation C

TABLE-US-00011 (a) Lactobacillus strain(s) 1 × 10.sup.10 CFU (b) Macrogol 4000 BP 350 mg

[0094] 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.

[0095] Formulation D (Controlled Release Capsule) 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-00012 (a) Lactobacillus strain(s) 1 × 10.sup.10 CFU (b) Microcrystalline Cellulose 125 mg (c) Lactose BP 125 mg (d) Ethyl Cellulose 13 mg

[0096] Example D: Powder Formulations

[0097] Formulation A (fast-melting microbial composition)

TABLE-US-00013 (a) Lactobacillus strain(s) 80 mg (preferably 1 × 10.sup.10 CFU) (b) Erythritol 450 mg (c) Inulin 227.5 mg (d) Xylitol 227.5 mg (e) Lemon flavour 10 mg (f) Silicon dioxide 5 mg

[0098] Formulation B (fast-melting microbial composition)

TABLE-US-00014 (a) Lactobacillus strain(s) 80 mg (preferably 1 × 10.sup.10 CFU) (b) Erythritol 425 mg (c) Inulin 215 mg (d) Xylitol 215 mg (e) Maltodextrin 50 mg (f) Lemon flavour 10 mg (g) Silicon dioxide 5 mg

[0099] Study of Effect of Lactobacillus Treatment on Mood States in Humans

[0100] The study was a randomized, placebo controlled, double-blind, parallel group, single centre study. The Wilcoxon signed rank test was used when evaluating change over time within each group and Wilcoxon rank sum test was used when evaluating differences between groups. All reported p-values are two-sided and nominal, i.e. not adjusted for multiple testing.

[0101] As shown in Table 2, the evaluation of mood states using the POMS questionnaire revealed increased vigour after the 12 weeks of intake of the Lactobacillus strain as compared to control (3.5 vs. 0.1, p=0.0151, change from baseline to 12 weeks).

[0102] Study Design

[0103] The study was initiated by the screening of healthy males and females at 16 to 40 years of age involved in recreational training or elite sports with a minimum of 5 hours training per week (Visit 1). A total of 365 subjects were screened. Subjects displaying iron deficiency (low level of serum ferritin, i.e. <30 μg/L) but without anemia (low level of Hb) were eligible to be randomized into the intervention which included 12 weeks (Visits 2-5) of daily administration of the investigational Lactobacillus strain-containing product or control product which did not contain the Lactobacillus strain.

[0104] Investigational Product

[0105] Capsules containing freeze-dried Lactobacillus plantarum DSM 9843 (registered trade marks LP299v®; 299v®) at a concentration of 1×10.sup.10 CFU/capsule, 20 mg iron (ferrous-fumarate), maize starch (bulking agent), maltodextrin (bulking agent), cellulose derivatives (coating of minerals) and magnesium stearate (processing aid/anti-caking agent). The capsules themselves consisted of hydroxypropylmethyl cellulose and titanium dioxide.

[0106] The control product contained all but the Lactobacillus strain. The investigational and control products were packed in blister packs with 10 capsules per blister. One box contained four blisters (40 capsules) to provide adequate amount for a 4-week administration period.

[0107] Each subject filled out a POMS questionnaire related to perceived mood state.

[0108] Subject Disposition

[0109] In total, 365 subjects were included in the study. After the screening and initial blood analysis, 53 subjects fulfilled all inclusion criteria and none of the exclusion criteria and were randomized. During the study, 14 subjects were excluded or withdrew their consent.

[0110] Baseline Characteristics

[0111] No eligible male subjects were found in the screening process and therefore only female subjects were included in the study. The subjects in the Lp299v group trained for an average of 8.2±2.0 hours per week before the start of the study and the subjects in the control group trained for an average of 7.6±2.0 hours per week. There were no differences in average training, age, body weight or BMI between the groups at the start of the intervention.

[0112] RESULTS

[0113] Profile of Mood Score Reveals Increased Vigour Upon Treatment With Lactobacillus

[0114] The Profile of Mood States (POMS) questionnaire is a validated psychological test developed by McNair et al. (McNair, D.; Lorr, M.; Doppleman, L. (1971). POMS Manual for the Profile of Mood States. San Diego, Calif.: Educational and Industrial Testing Service). It was used to obtain a Total Mood Disturbance score and to analyse the Tension,

[0115] Depression, Anger, Vigour, Fatigue and Confusion sub-scores (see Table 2).

TABLE-US-00015 TABLE 2 Results from the POMS questionnaire at baseline and after 4, 8 and 12 weeks of supplementation. Baseline 4 weeks 8 weeks 12 weeks ΔV3-V2 mean std mean std mean std mean std mean Std (p-value) Total Control 124 22 120 20 122 34 118 29 −3.2 23.2 (0.9128) POMS Lp299v 129 30 126 30 120 25 120 27 −3.3 19.4 (0.9128) Vigor Control 58 8 58 9 58 11 59 7 −0.2 6.5 (0.6232) Lp299v 58 6 60 6 60 8 61 6 1.4 4.9 (0.6232) Tension Control 42 6 40 5 40 7 40 8 −1.9 6.3 (0.9213) Lp299v 43 6 41 4 40 5 41 6 −1.8 3.9 (0.9213) Depression Control 40 5 38 4 39 6 38 6 −0.3 5.3 (0.2662) Lp299v 41 6 41 8 40 5 40 7 0.5 5.2 (0.2662) Anger Control 44 6 43 5 46 9 43 6 −0.05 5.8 (0.3720) Lp299v 45 9 44 7 44 6 44 5 −1.2 4.9 (0.3720) Fatigue Control 49 7 49 6 48 9 47 8 0.7 6.4 (0.9659) Lp299v 48 8 49 9 47 8 48 7 1 6.8 (0.9659) Confusion Control 41 5 41 5 40 7 40 6 −0.6 5.0 (0.4689) Lp299v 43 5 42 6 40 4 42 6 −1.4 4.4 (0.4689) n Control 23 NA 22 NA 15 NA 16 NA 22 NA Lp299v 17 NA 17 NA 16 NA 16 NA 16 NA ΔV4-V2 ΔV5-V2 mean Std (p-value) mean std (p-value) Total Control 4.3 32.3 (0.2994) 4.6 22.4 (0.2199) POMS Lp299v −11 30.2 (0.2994) −12.6 42.1 (0.2199) Vigor Control 1.2 9.8 (0.6894) 0.13 3.5 (0.0151) Lp299v 2.6 8.9 (0.6894) 3.5 6.3 (0.0151) Tension Control −0.9 5.8 (0.5046) −0.46 5.7 (0.5179) Lp299v −2.7 5.5 (0.5046) −3.2 8.3 (0.5179) Depression Control 0.7 0.67 (0.9754) 1.7 5.5 (1.000) Lp299v −1.2 6.1 (0.9754) −1.15 6.6 (1.000) Anger Control 4.1 6.9 (0.0934) 2.3 5.4 (0.5012 Lp299v −0.7 7.8 (0.0934) −1.6 11.4 (0.5012) Fatigue Control 1.3 8.3 (0.6490) 1.9 6.8 (0.4116) Lp299v −2 8.2 (0.6490) −2.6 10.9 (0.4116) Confusion Control 0.07 6.2 (0.2602) 0.5 4.6 (0.5664) Lp299v −3.3 5.7 (0.2602) −2.34 7.6 (0.5664) n Control 15 NA 16 NA Lp299v 15 NA 15 NA V2 = Baseline V3 = 4 weeks of supplementation V4 = 8 weeks of supplementation V5 = 12 weeks of supplementation

[0116] The results from the questionnaire show that the sub-score Vigour increased after 12 weeks of supplementation in the Lactobacillus treatment group (from a mean 58.0 at baseline to 61.43 at week 12, n=15-17, change=3.53, n=15, p=0.0436), but no differences were detected in the control group (from a mean 58.0 to 59.4, n=16-23, change=0.13, n=16, p=0.9814).

[0117] Also, the change from baseline to week 12 was significantly different compared to the control group (3.5 vs.0.13, p=0.0151).

[0118] No other differences between the groups could be detected neither for the total profile (from a mean 128.8 at baseline to 120.0 for Lp299v and 123.9 to 118 for control, change=−12.6 vs. 4.56, p=0.2199 nor for any of the other sub-scores (see Table 2).

[0119] Conclusions

[0120] The above study in humans shows that treatment by Lactobacillus plantarum administration resulted in an increase in vigour.

[0121] The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.