WO2022245882A1

WO2022245882A1 - Supplements for supporting gastrointestinal health

Info

Publication number: WO2022245882A1
Application number: PCT/US2022/029711
Authority: WO
Inventors: Guruduth S. Banavar; Hilary KEISER; Joshua Grant ANTOINE; Janelle CONNELL
Original assignee: Viome Life Sciences, Inc.
Priority date: 2021-05-17
Filing date: 2022-05-17
Publication date: 2022-11-24
Also published as: WO2022245849A2; WO2022245875A1; WO2022245849A3

Abstract

Provided here are compositions that support gastrointestinal health. The compositions comprise one or more a mixture of heat-killed Lactobacillus fermentum and Lactobacillus delbruceckii; an amylase, a cellulase, a lactase, a lipase, a protease, bromelain, horse chestnut seed extract, peppermint leaf extract, deglycyrrhizinated licorice root extract, vitamin A, vitamin C, pectin (e.g., apple pectin), butyrate, ginger root extract, and resveratrol.

Description

Supplements for Supporting Gastrointestinal Health

REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of the priority date of U.S. provisional application 63/189,566, filed May 17, 2021 , the contents of which are incorporated herein in their entirety.

BACKGROUND

[0002] Gastrointestinal health is important, not only for nutrient absorption, but also for immune function and mental health. Gastrointestinal discomforts such as, for example, abdominal pain, constipation, diarrhea, cramping, bloating and gas are experienced by many individuals and, in addition to discomfort, can lead to lost productivity. Current treatments for gastrointestinal discomfort often comprise synthetic materials such as bismuth compounds and are often ineffective.

SUMMARY

[0003] Provided herein are supplements for supporting or improving gastrointestinal health in a subject (e.g., an animal, a mammal, a primate or a human) made from natural ingredients. Compositions disclosed herein are formulated to reduce symptoms of poor gastrointestinal health such as, for example, abdominal pain, constipation, diarrhea, cramping, bloating and gas. The compositions and formulations disclosed herein target and, optionally, modulate, microbial pathways (e.g., synthetic pathways and/or degradative pathways) that are dysregulated in humans with poor gastrointestinal health. Each formula ingredient targets one or more of these dysregulated pathways; the outcome being the synergy of all ingredients to effectively influence these microbial pathways to improve gastrointestinal health, correct abnormalities in the gastrointestinal tract, and/or relieve symptoms of gastrointestinal discomfort. Thus, for example, administration of the compositions and/or formulations described herein can help ameliorate, e.g., abdominal pain, constipation, diarrhea, cramping, bloating and gas; and lead to improved nutrient uptake, improved immune function, and improved general health.

[0004] The compositions comprise Lactobacillus Postbiotic, as described herein. In certain embodiments, the compositions comprise Lactobacillus Postbiotic and any three of an amylase, a cellulase, a lactase, a lipase, a protease, and bromelain.

[0005] In additional embodiments, the compositions comprise Lactobacillus Postbiotic and any four of an amylase, a cellulase, a lactase, a lipase, a protease, and bromelain.

[0006] In further embodiments, the compositions comprise Lactobacillus Postbiotic and any five of an amylase, a cellulase, a lactase, a lipase, a protease, and bromelain. [0007] In additional embodiments, the compositions comprise Lactobacillus Postbiotic and all six of an amylase, a cellulase, a lactase, a lipase, a protease, and bromelain.

[0008] Exemplary amylases include alpha-amylase, beta-amylase, gamma-amylase, and maltase.

[0009] Exemplary cellulases include endo-1 ,4-beta-D-glucanase, carboxymethyl cellulase, avicelase, celludextrinase, cellulase A, cellulosin AP, alkali cellulase, cellulase A 3,

9.5 cellulase, and pancellase SS.

[0010] Exemplary lactases include lactase-phlorizin hydrolase (aka LPH).

[0011] Exemplary lipases include pancreatic lipase isoforms L1 and L3.

[0012] Exemplary proteases include neutral protease, acid protease, alkaline protease, chymotrypsin, ficin, papain, serrapeptase, and trypsin.

[0013] In additional embodiments, the compositions comprise Lactobacillus Postbiotic and any one of horse chestnut seed extract, peppermint leaf extract and deglycyrrhizinated licorice.

[0014] In additional embodiments, the compositions comprise Lactobacillus Postbiotic and any two of horse chestnut seed extract, peppermint leaf extract and deglycyrrhizinated licorice.

[0015] In additional embodiments, the compositions comprise Lactobacillus Postbiotic and all three of horse chestnut seed extract, peppermint leaf extract and deglycyrrhizinated licorice.

[0016] In further embodiments, the compositions comprise:

(a) Lactobacillus Postbiotic,

(b) any three, any four, any five, or all six of an amylase, a cellulase, a lactase, a lipase, a protease, and bromelain, and

(c) any one, any two or all three of horse chestnut seed extract, peppermint leaf extract and deglycyrrhizinated licorice.

[0017] In still further embodiments, any of the foregoing compositions can also include vitamin A, vitamin C, or both vitamin A and vitamin C.

[0018] In additional embodiments, any of the foregoing compositions can also include pectin (e.g., apple pectin), butyrate, ginger root extract and/or resveratrol.

[0019] In certain embodiments, the amount of heat-killed L fermentum and L delbruceckii cells in the Lactobacillus Postbiotic is at least 1 ^c 10^s, 1 ^c 10⁷, 1 ^c 10^s, 1 ^c 10⁹, 1 ^c 10¹⁰, 1 x 10¹¹ , or 1 X 10¹² colony-forming units (CFU). In additional embodiments, the amount of heat-killed L. fermentum and L. delbruceckii cells in the Lactobacillus Postbiotic is between 1 x 10⁸ and 1 x 10¹⁴ CFU, between 1 10⁸ and 1 x 10¹³ CFU, between 1 x 10⁹ and 1 x 10¹³ CFU or between 1 x 10^® and 1 x 10¹² CFU. In further embodiments, the amount of heat-killed L fermentum and L delbruceckii cells in the Lactobacillus Postbiotic is between 1 x 10¹⁰ and 3 x 10¹⁰ CFU. In another embodiment, the amount of heat-killed L fermentum and L delbruceckii cells in the Lactobacillus Postbiotic is 2 x 10¹⁰ CFU.

[0020] The compositions can be provided as dosage forms, for example, a liquid, a capsule, a tablet, a gummy, a stick pack (e.g., pixie stick), a blister pack, and a packet. A daily dose can comprise one, two, three, four or more dosage forms. A daily dose can comprise a reference daily intake (“RDI”), a recommended daily allowance or an expert-determined dose for each of a plurality of, e.g., all or a majority of ingredients in the daily dose. For example, the reference daily intake can be an amount recommended by a government authority, such as the Food and Drug Administration, or recommended by a non-government authority or a healthcare provider such as a doctor, nurse practitioner, or nurse.

BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The accompanying drawings, which are incorporated herein and form a part of the specification, illustrate exemplary embodiments and, together with the description, further serve to enable a person skilled in the pertinent art to make and use these embodiments and others that will be apparent to those skilled in the art.

[0022] FIG. 1 shows an exemplary composition for supporting gastrointestinal health:

“gastrointestinal Health Formula 3.2”.

[0023] FIG. 2 shows an exemplary composition for supporting gastrointestinal health:

“gastrointestinal Health Formula 3.4”.

[0024] FIG. 3 shows exemplary compositions for supporting gastrointestinal health:

“gastrointestinal Health Formula 4.2”, and “gastrointestinal Health Formula 4.4”.

DETAILED DESCRIPTION

I. Introduction

[0025] A systems biology platform that incorporates high-resolution molecular data generated from biological specimens, diet information, accurate clinical data, domain knowledge of microbial and human physiology, and data science/AI was used for data analysis. Data (e.g., from samples of stool, blood, urine, sputum, saliva) obtained from over 10,000 subjects was analyzed, using the above-mentioned systems biology platform, to identify gut microorganisms and biochemical pathways related to gastrointestinal health. The compositions described herein modulate growth of gut microbes and/or modulate microbial biochemical pathways, to improve gastrointestinal health and reduce symptoms of gastrointestinal discomfort.

[0026] Gastrointestinal discomfort is associated, in some circumstances, with specific alterations in gut microbial activity. These activities are due to a combination of an individual’s microbiome composition (e.g., taxonomy) and diet. Observational studies have identified specific biochemical pathways in the gut microbiome that are associated with gastrointestinal discomfort. Based on the data and models developed from the observational studies described above, the compositions disclosed herein were developed to modulate the molecular ingredients in the diet to inhibit microbial functions that trigger and exacerbate gastrointestinal dysfunction, and to activate microbial pathways associated with gastrointestinal health.

[0027] For gastrointestinal health, a combination of unique supplements and their amounts were designed using domain knowledge-based interpretations and machine-learned models from the cross-sectional and longitudinal molecular and symptoms data from more than 100,000 people.

[0028] Supplement compositions are pre-formulated into products designed to improve gastrointestinal health in subjects. These supplements can be purchased in stores or delivered to a subject’s home.

[0029] Compositions are designed to rebalance the microbial composition and overall microbiome functional activities, e.g., biochemical pathways, of each subject. This can be performed at the specific molecular pathway level (e.g., boosting the activity levels of butyrate production pathways) or at the integrative functional category or profile level (e.g., improving gastrointestinal health and inflammatory activity functional patterns).

[0030] Learning algorithms implemented on data from subjects scoring positively on a clinically validated questionnaire for irritable bowel disease identified biochemical pathways and microbial taxa associated with the conditions. The ingredients selected here affect or regulate one or more of the associated microbial taxa and biochemical pathways.

[0031] A strong correlation was observed between gastrointestinal discomfort and low expression of gut microbial pathways related to food digestion in the gut microbiome. Accordingly, certain compositions disclosed herein address the digestive efficiency of the gut microbiome.

[0032] A strong correlation was observed between gastrointestinal discomfort and high expression levels of gut microbial pathways that result in protein fermentation. Therefore, certain compositions comprise ingredients that ameliorate protein fermentation and its byproducts.

[0033] A strong correlation was observed between subjects exhibiting gastrointestinal discomfort and high expression levels of inflammatory gut microbial pathways. Accordingly, certain compositions disclosed herein address high microbiome inflammatory activity by reducing inflammation in the body.

[0034] A strong correlation was observed between subjects exhibiting gastrointestinal discomfort and high prevalence of viruses in the gut microbiome. Therefore, certain compositions comprise ingredients that have antiviral activity.

[0035] A strong correlation was observed between subjects exhibiting gastrointestinal discomfort and low active gut microbial diversity. Therefore, certain compositions comprise ingredients that encourage diversity in the microbiome.

[0036] A strong correlation was observed between gastrointestinal discomfort and high expression levels of gut microbial LPS (lipopolysaccharide) production pathways. Therefore, certain compositions comprise ingredients that reduce LPS production.

[0037] A strong correlation was observed between gastrointestinal discomfort and high expression levels of gut microbial methane gas producing pathways. Therefore, certain compositions comprise ingredients that reduce methane gas production. [0038] A strong correlation was observed between gastrointestinal discomfort and high expression levels of gut microbial biofilm formation pathways. Therefore, certain compositions comprise ingredients that reduce biofilm formation.

[0039] There was a strong correlation between subjects exhibiting gastrointestinal discomfort and high expression levels of gut microbial pathways that result in the production of flagella (an appendage that enables motility and adherence to the intestinal lining). Therefore, certain compositions comprise ingredients that reduce motility and adherence and/or reduce the production of flagella by bacteria.

[0040] There was a strong correlation between subjects exhibiting gastrointestinal discomfort and low abundance of butyrate producing taxa. Therefore, certain compositions comprise ingredients that stimulate butyrate production pathways and/or enhance the growth of butyrate- producing gut bacteria. In addition, certain compositions disclosed herein comprise butyrate. Ingredients for Supporting gastrointestinal health

[0041] The following ingredients support gastrointestinal health at the microbiome or the condition levels. They can be included in varying combinations and relative amounts in supplement compositions for supporting gastrointestinal health.

[0042] FIGs 1 and 2 show specifications identifying compounds in the respective ingredients that contribute to the function of the ingredient in the supplement composition. In any supplement combination described herein, the ingredient can be substituted with the specified compound to obtain the desired result. A. Amylases

1. Microbiome: a) Low digestive efficiency pathways

[0043] Amylases are enzymes that break down carbohydrates. Accordingly, ingestion of amylase can improve efficiency of digestion. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC6825871/.

B. Cellulases

1. Microbiome: a) Low digestive efficiency pathways

[0044] Cellulases are, as the name suggests, enzymes that break down cellulose.

Accordingly, ingestion of a cellulase can increase digestive efficiency. See, e.g., www.intechopen.com/chapters/66517.

C. Lactases

1. Microbiome: a) Low digestive efficiency pathways

[0045] Lactases are enzymes that break down lactose. Accordingly, ingestion of a lactase can increase digestive efficiency, especially for those with diets high in dairy products or those who are lactose-intolerant. (See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC6669050/.

2. Gastrointestinal Health Support

[0046] Low levels of lactase have been associated with inflammatory bowel disease

(IBD). See, e.g., nutritionj. biomedcentral.com/articles/10.1186/s12937-016-0183-8.

D. Lipases

1. Microbiome: a) Low digestive efficiency pathways

[0047] Lipases are enzymes that digest lipids; i.e., they break down fat. Accordingly, ingestion of a lipase can improve digestive efficiency. See, e.g., journals.lww.com/pancreasjournal/Abstract/2003/01000/Lipase_Supplementation_Therapy _ St andards,.1.aspx

E. Proteases

1. Microbiome: a) Low digestive efficiency pathways [0048] Proteases are enzymes that hydrolyze peptide bonds in proteins and peptides. www.ncbi.nlm.nih.gov/pmc/articles/PMC4030975/. Some proteases cleave the a-peptide bonds between amino acids in naturally-occurring proteins. www.ncbi.nlm.nih.gov/pmc/articles/PMC2576539/ Accordingly, proteases can improve digestive efficiency by helping to break down proteins in food.

F. Bromelain

[0049] Bromelain's preferential cleavage site is the carbonyl end of lysine, alanine, tyrosine and glycine.

[0050] It has been demonstrated that bromelain's specificity is similar to that of the endogenous protease plasmin. Bromelain acts on fibrinogen to give products that are similar, at least in effect, to those formed by plasmin. They are small molecular weight active peptides, which regulate prostaglandin biosynthesis and create conditions existing in the healthy organism. See, e.g., www.sciencedirect.com/science/article/abs/pii/0306987780900389#:~:text=Bromelain%20acts%

20on%20fibrinogen%20to,existing%20in%20the%20healthy%20organism.

1. Microbiome: a) Low digestive efficiency

[0051] Bromelain is a protease that is derived from pineapple. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC3529416/.

2. Protein fermentation

[0052] Bromelain possesses proteolytic activity (breaks down proteins), thereby assisting in digestion.

[0053] www.ncbi.nlm.nih.gov/pmc/articles/PMC3529416/

3. Inflammatory activity

[0054] Treatment of cells with bromelain inhibits activation of CD4⁺ T-cells and reduces expression of CD25 by T-cells. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC3529416/.

4. LPS biosynthesis

[0055] Bromelain has been shown to inhibit induction of NF-KB activity by bacterial endotoxin (LPS). Bromelain also inhibits expression of prostaglandin E2 (PGE2) and cyclooxygenase 2 (Cox-2) in human monocytic leukemia cell lines and in murine microglial cell lines. See, e.g., http://www.tandfonline.com/doi/full/10.1080/08820130802083622. a) Gastrointestinal Health Support: [0056] Bromelain supplementation reduces expression of pro-inflammatory markers in cells. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC4998156/; www.ncbi.nlm.nih.gov/pubmed/15936249. Bromelain has been shown to improve digestive efficiency See, e.g., www.jstage.jst.go.jp/article/jphs1951/22/4/22_4_519/_article.

G. Horse chestnut seed extract

[0057] In certain embodiments, a methanol extract of horse chestnut seeds is used.

Seeds are pulverized and extracted with methanol. The methanol phase is then dried and the residue is powerdized.

1. Microbiome: a) Methane Production Pathways

[0058] Horse chestnut extract reduces the amount of methanogenic archaebacteria in the small intestine, thereby also reducing the amount of methane in the small intestine. Tannins produced by the horse chestnut seed, and present in its extract, act as molecular sponges to adsorb hydrogen. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC4986399/.

2. Gastrointestinal Health Support

[0059] Horse chestnut has been used to treat IBS-C (the constipation dominant variety of irritable bowel syndrome). See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC4986399/.

H. Peppermint leaf extract

[0060] In certain embodiments, a hot water extract of peppermint leaves is used.

Leaves are pulverized and extracted with hot water. The aqueous extract is then dried, and the powder is used in the compositions disclosed herein.

1. Gastrointestinal Health Support:

[0061] Peppermint has recognized antispasmodic effects. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC6337770/.

I. Lactobacillus Postbiotic

[0062] “Lactobacillus Postbiotic” refers to a fermentate and microbial biomass generated by heat-treating two Lactobacillus strains, Lactobacillus fermentum, and Lactobacillus delbrueckii. Lactobacillus Postbiotic can be made by growing a mixed culture of mixture of Lactobacillus fermentum and Lactobacillus delbruceckii to a particular cell density (specified elsewhere herein) and, when the desired cell density is reached, subjecting the culture to heat inactivation to kill the cells. The heat-inactivated culture comprises dead cells, and metabolites produced by the cells. One version of Lactobacillus Postbiotic is available from Adare Pharmaceuticals, Houdain, France as LBiome(™). See, e.g., WO 2019/180263.

1. Microbiome: a) Inflammatory Activity

[0063] Lactobacillus Postbiotic administration to mice reduced pathogen-induced inflammation. (See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC7003559/) b) Flagellar Assembly Pathways

[0064] Lactobacillus Postbiotic reduced swimming motility of pathogen S. enterica serovarTyphimurium within cultured human enterocyte-like cells. (See, e.g., journals.asm.org/doi/full/10.1128/AAC.00418-11.) c) Intestinal Barrier Health

[0065] Lactobacillus Postbiotic inhibits adhesion of enteroinvasive pathogens to human intestinal Caco-2 cells by and decreases bacterial invasion.

(See, e.g., pubmed.ncbi.nlm.nih.gov/8354463/; pubmed.ncbi.nlm.nih. gov/1624102/; www.jstor.org/stable/23498285; pubmed.ncbi.nlm.nih. gov/1527509/.)

2. Gastrointestinal Health Support

[0066] Lactobacillus Postbiotic improves gastrointestinal function, reduces inflammation, and reduces symptoms of diarrhea.

J. Deglycyrrhizinated Licorice (DGL) Root Extract

1. Microbiome:

[0067] In certain embodiments, an acetone extract of licorice roots is used. Roots are pulverized and extracted with acetone. The acetone phase is then dried and the powder is used in the formulations described herein. a) Intestinal barrier health

[0068] In human trials, DGL has been found to support a healthy gastric mucosal lining by stimulating mucus production and enhancing blood supply to the intestinal mucosa. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC3123991/. b) Inflammatory Activity

[0069] Licorice reduces gastric inflammation by inhibiting prostaglandin synthesis and inhibiting production of lipoxygenase. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC4928719/.

2. Gastrointestinal Health Support

[0070] DGL is commonly used for treatment of gastrointestinal disorders such as ulcerative colitis, peptic ulcers, and canker sores; and is used to treat inflammatory bowel disease. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC3123991/.

K. Vitamin A

1. Microbiome:

[0071] Vitamin A deficiency damages both humoral and cellular immunity in the intestinal mucosa. See, e.g., pubmed.ncbi.nlm.nih.gov/20363594/.

L. Vitamin C

1. Microbiome: a) Intestinal Barrier Health

[0072] Vitamin C improves intestinal barrier function by upregulating collagen synthesis by in the intestine. See, e.g., www.ncbi.nlm.nih.gov/pmc/articles/PMC6327911/. b) Biofilms

[0073] Vitamin C reduces synthesis of microbial extracellular polymeric substances

(e.g., biofilms) by S. epidermidis, E. coli, B. subtilis, and P. aeruginosa in in-vitro studies. www.frontiersin.org/articles/10.3389/fmicb.2017.02599/full?report=reader. Biofilms are used as protective mechanisms by certain microbes. c) Antiviral activity

[0074] Vitamin C has demonstrated antiviral activity. See, e.g., www.spandidos- publications.com/ijmm/22/4/541.

M. Pectin

[0075] Pectin (e.g., from apples, berry, and citrus fruits) has a number of beneficial effects on the digestive system. 1. Microbiome a) Microbial diversity

[0076] Apple pectin supplementation reverses detrimental changes in the microbiome that result from a high-fat diet. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC4808856/. b) Butyrate production

[0077] Bacterial decomposition of pectin produces short-chain fatty acids. See, for example, pubmed.ncbi.nlm.nih.gov/16889120/. c) Intestinal barrier Health

[0078] Apple pectin has been shown to improve intestinal barrier function. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC4808856/. d) Digestive efficiency

[0079] Pectin enhances the intestinal solubility and absorption of ferric iron. See, for example, pubmed.ncbi.nlm.nih.gov/24864109/.

2. Gastrointestinal Health Support

[0080] Pectin acts as a prebiotic in specifically stimulating gut bifidobacteria in IBS-D patients and is effective in alleviating clinical symptoms, balancing colonic microflora and relieving systemic inflammation. See, for example, pubmed.ncbi.nlm.nih.gov/25809332/

N. Butyrate

1. Microbiome a) Inflammatory Activity

[0081] Butyrate is capable of reducing inflammation by inhibiting the growth of pathobionts and by increasing the integrity of the mucosal barrier of the small intestine [CORRECT?]. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC7192831/. b) Butyrate producer taxa

[0082] Supplementation with butyrate can be used if butyrate-producing organisms are not present in the gastrointestinal system of the subject. c) Intestinal barrier health

[0083] Butyrate stimulates production of MUC2 mucin in a human colonocyte cell line, which helps reinforce the colonic defense barrier. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC3070119/. 2. Gastrointestinal Health Support

[0084] Butyrate has been used for the treatment of inflammatory bowel disease (IBD) and Crohn’s disease. See, for example, pubmed.ncbi.nlm.nih.gov/30277149/ and www.sciencedirect.com/science/article/abs/pii/S1594580408600091 .

O. Ginger root extract

1. Microbiome

[0085] Ginger root extract has been used traditionally to reduce gas and bloating. See, for example, pubmed.ncbi.nlm.nih.gov/23612703/.

2. Gastrointestinal Health Support

[0086] Ginger root extract has been used traditionally to reduce gas and bloating. See, for example, pubmed.ncbi.nlm.nih.gov/23612703/.

P. Resveratrol

1. Microbiome a) Inflammatory activity

[0087] Resveratrol has been shown to prevent and ameliorate intestinal inflammation.

See, for example, pubmed.ncbi.nlm.nih.gov/29191255/. b) Flagellar assembly and biofilm formation

[0088] Resveratrol has been shown to reduce swarming motility of, and biofilm formation by, E. coli. See, for example, pubmed.ncbi.nlm.nih.gov/24067082/.

2. Gastrointestinal Health Support

[0089] Administration of resveratrol to individuals suffering from ulcerative colitis reduced plasma levels of tumor necrosis factor alpha (TNF-a) and C-reactive protein (CRP). See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC4220497/.

Q. L. Plantarum 299v

1. Microbiome a) Ammonia production

[0090] L. plantarum directly consumes ammonia in the gut, according to in-vivo models

See, for example, pubmed.ncbi.nlm.nih.gov/18697211/; pubmed.ncbi.nlm.nih.gov/11711768/; pubmed.ncbi.nlm.nih. gov/10811333/. 2. Gastrointestinal Health Support

[0091] In several studies, L. plantarum 299v has demonstrated ability to improve signs and symptoms of IBS such as bloating, abdominal pain, and flatulence. See, for example, www.ncbi.nlm.nih.gov/pmc/articles/PMC3419998/.

II. Compositions for Supporting Gastrointestinal Health

[0092] In some embodiments, the ingredients in a composition comprises, consists essentially of, or consists of at least any of two, three, four, five, six, seven, eight, nine, ten or eleven, twelve, thirteen, fourteen, fifteen or sixteen ingredients selected from:

• Amylase (e.g., 5,000 DU to 7,000 DU);

• Cellulase (e.g., 25 to 500 CU);

• Lactase (e.g., 750 ALU to 1250 ALU);

• Lipase (e.g., 20 FlP to 70 FIP);

• Protease (e.g., 1250 PC to 1750 PC);

• Bromelain (e.g., 10 GDU to 100 GDU );

• Apple pectin (e.g., 200 mg to 600 mg);

• Butyrate (e.g., 100 mg to 500 mg);

• Ginger Root Extract (e.g., 25 mg to 700 mg);

• Resveratrol (e.g., 25 mg to 500 mg);

• Horse Chestnut Seed Extract (e.g., 50 mg to 700 mg);

• Peppermint Leaf Extract (e.g., 100 mg to 700 mg);

• Deglycyrrhizinated Licorice (DGL) Root Extract (e.g., 26 mg to 500 mg);

• Vitamin A (e.g., 0.5 mg to 5 mg);

• Vitamin C (e.g., 20 mg to 750 mg); and

• Lactobacillus Postbiotic (e.g., 10 B CFU to 30 B CFU.

[0093] In another embodiment, the composition comprises, consists essentially of, or consists of:

• One, two, three, four, five or six of: an amylase, a cellulase, a lactase, a lipase, a protease and bromelain;

• One, two or three of: a horse chestnut seed extract, a peppermint leaf extract and deglycyrrhizinated licorice root extract; and

• Lactobacillus Probiotic.

[0094] The composition can further comprise, consist essentially of, or consist of one, two, three, four, five or six of:

• Vitamin A;

• Vitamin C;

• Pectin; • Butyrate;

• Ginger root extract; and

• Resveratrol.

[0095] In certain embodiments, the composition comprises Lactobacillus Postbiotic, and three, four, five or six ingredients selected from an amylase, a cellulase, a lactase, a lipase, a protease or bromelain.

[0096] In one embodiment, the composition comprises:

• An enzyme blend comprising o an amylase, o a cellulase, o a lactase, o a lipase, and o a protease;

• Bromelain;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Deglycyrrhizinated Licorice (DGL) Root Extract; and

• L. plantarum 299v.

[0097] In one embodiment, the composition comprises:

• An enzyme blend comprising amylase, a cellulase, a lactase, a lipase, and a protease;

• Apple pectin;

• Bromelain;

• Butyrate;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Ginger Root Extract;

• Resveratrol;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin C; and

• L. plantarum 299v.

[0098] In one embodiment, the composition comprises:

• Bromelain;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Deglycyrrhizinated Licorice (DGL) Root Extract; • Vitamin A;

• Vitamin C; and

• Lactobacillus Postbiotic.

[0099] In one embodiment, the composition comprises:

• Bromelain;

• Apple pectin;

• Butyrate;

• Ginger Root Extract;

• Resveratrol;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin A;

• Vitamin C; and

• Lactobacillus Postbiotic.

[0100] In one embodiment, at least one ingredient of the composition is a probiotic or a postbiotic.

[0101] The composition of any of the preceding claims wherein the ingredients, together, improve at least any of at least any of one, two, three, four, five, six, seven, eight, nine, ten or eleven of the following: low digestive efficiency, high inflammatory activity, high bacterial LPS biosynthesis, increased expression of flagellar assembly pathways, low digestive efficiency pathways, high protein fermentation pathways, low intestinal barrier health pathways, high biofilm formation pathways, high prevalence of virus taxa, low microbial diversity, and low numbers of butyrate producers.

[0102] In certain embodiments, a composition comprises a plurality of ingredients, which ingredients, together, provide gastrointestinal health support and microbiome support. In certain embodiments, the composition comprises ingredients that, together, improve at least any of one, two, three, four, five or six, of the following: abdominal pain, constipation, diarrhea, cramping, bloating and gas.

[0103] In certain embodiments, ingredients are present in relative amounts such that a single dose of the composition comprises about a recommended daily intake of each of the ingredients. Dosages found to be effective in clinical trials were used as a benchmark for amounts included in the dosages used here.

[0104] A dosage form can consist essentially of the named ingredients. Ingredients that do not materially affect the basic and novel characteristics of a claimed composition include, for example, excipients (liquids or solids), fillers, binders, capsules, coloring agents, flavoring agents and the like.

[0105] The composition can further comprise a pharmaceutically acceptable excipient, that is, a component other than an active ingredient that is safe and non-toxic for administration. Excipients typically function as carriers for active ingredients. Pharmaceutically acceptable excipients include, without limitation, water, saline, powders, pastes, creams, gels, starches, gums and oils.

[0106] In some embodiments, the composition can comprise or consist essentially of one or more ingredients from Table 1 and a pharmaceutically acceptable excipient and/or a coloring or flavoring agent.

A. Doses and Dosage Forms of Compositions for Supporting Gastrointestinal Health

[0107] A dose of a composition is an amount of the composition to be taken at one time.

A dosage form is the physical form of a composition to be administered to a subject. Dosage is the administration of a predetermined amount, number, and/or frequency of doses of a composition over a specific period of time. Accordingly, a dose of a composition can be contained in one or a plurality of dosage forms. For example, a dose of a composition as disclosed herein can be comprised in two, three, four, five or six dosage forms, e.g., tablets. All the ingredients in a dose of the composition can be in a single dosage form. Alternatively, a plurality of dosage forms can contain all the ingredients in a dose of the composition, e.g., with not all of the ingredients present in a single dosage form. a) Doses

[0108] Doses of the compositions described herein comprise specified amounts of the recited ingredients. These amounts can be effective to promote positive gastrointestinal health when taken regularly, for example, daily.

[0109] The amount of any ingredient in a dose can be a reference daily intake (“RDI”), a recommended daily allowance or an expert-determined dose for the ingredient. For example, the reference daily intake can be an amount recommended by a government authority, such as the Food and Drug Administration, or recommended by a non-government authority or a healthcare provider such as a doctor or nurse. Exemplary doses are set forth in FIGs. 1-3. Doses also can include one or a plurality of the following ingredients in amounts or about the amounts given. Accordingly, the amounts of each ingredient can constitute a dose of the ingredient to be administered at one time:

A) TABLE 1:

Key: DU = dextrinizing units CU = cellulase units ALU = acid lactase units FIP = Federation International pharmaceutical units

PC = protease casein units GDU = gelatin digestion units CFU = colony-forming units b) Dosage Forms

[0110] A dosage form for a composition of this disclosure can be any form typically consumed by a subject, e.g., a human. Many dosage forms are known in the art. Useful dosage forms include, for example, a liquid (e.g. , a syrup) a capsule, a tablet, a gummy, a stick pack (e.g., pixie stick), a blister pack and a packet (e.g., a tear packet). Capsules come in various sizes. These include, for example, #000 (1 .37 ml), #00 (0.93 ml), #0 (0.68 ml), #1 (0.50 ml), #2 (0.37 ml), #3 (0.30 ml) and #4 (0.21 ml). Packets may be foil packets. They may be tearable to make opening easier. The contents can be poured out, for example, into a beverage, for consumption.

[0111] A dose of a composition of this disclosure can be comprised in a single dosage form (e.g., one capsule) or a plurality of dosage forms (e.g., 2, 3, 4, 5 or 6 capsules). A single dosage form can include all of the ingredients in a composition. Alternatively, the ingredients in a single dose can be divided between a plurality of dosage forms.

B) Methods of Supporting gastrointestinal health

[0112] In another aspect provided herein is a method for supporting gastrointestinal health, comprising administering an effective amount of a composition as disclosed herein to a subject in need thereof. For example, the composition can be administered as a dosage form orally. a) Dosages

[0113] A dosage can comprise an effective amount of composition to improve one or more signs or symptoms of gastrointestinal discomfort. Such signs or symptoms include, for example, at least abdominal pain, abdominal distension, constipation, diarrhea, cramping, bloating and gas.

[0114] Dosages of a composition of this disclosure for a subject, such as a human, can be a single dose, as described herein, provided daily or nearly daily over a period of time. For example, the dosage could be a single dose taken one, two, three, four, five, six or seven days a week, over the course of at least one week, at least one month, at least four months or at least one year.

C) Kits

[0115] In another aspect provided herein is a kit comprising a plurality of doses or daily dosage forms of any of the compositions described herein. For example, the kit can comprise a single container, such as a bottle, ajar, a vial, or a bag that contains a plurality of dosage forms. In another embodiment the kit comprises a blister pack that comprises a plurality of pockets, each of which comprises one or more dosage forms. [0116] A kit can further include a shipping container adapted to hold one or more containers, e.g., as described above, and adapted to transmit the containers to a recipient, such as a customer. The shipping container can be, for example, an envelope, a bag, a box or a shipping tube of sufficient volume to hold the one or more containers. The shipping containers can be adapted for transport to distant locations, for example locations that are at least any of 1 mile, 10 miles, 100 miles or 1000 miles away from the point of shipping or located in another city, state or country. The shipping container can be any container suitable for shipping through a common carrier. Common carriers include, for example, a national postal service (e.g., the United States Postal Service) and private companies (e.g., FedEx, DHL and UPS).

[0117] The kit can optionally include instructions for use thereof.

[0118] In another embodiment, the composition is formulated as a dose comprising one, two, three or four capsules.

EXEMPLARY EMBODIMENTS

[0119] 1. A composition comprising:

(a) a Lactobacillus Postbiotic;

(b) any three, four, five or six enzymes selected from:

(i) an amylase,

(ii) a cellulase,

(iii) a lactase,

(iv) a lipase,

(v) a protease, and

(vi) bromelain; and

(c) any one, two or three extracts selected from:

(i) horse chestnut seed extract,

(ii) peppermint leaf extract, and

(iii) deglycyrrhizinated licorice.

[0120] 2. The composition of embodiment 1 , comprising all six of the enzymes of (b).

[0121] 3. The composition of embodiment 1 or 2, comprising all three of the extracts of

(c).

[0122] 4. The composition of any of the preceding embodiments, further comprising one or both of: (a) vitamin A, and

(b) vitamin C.

[0123] 5. The composition of any of the preceding embodiments, further comprising one or more of:

(a) apple pectin;

(b) butyrate;

(c) ginger root extract; and

(d) resveratrol.

[0124] 6. The composition of any of the preceding embodiments, comprising enzymes:

Lactobacillus Postbiotic (gms:B CFU) of between about 5:1 and about 80:1 , e.g., about 5:1 .

[0125] 7. The composition of any of the preceding embodiments, comprising enzymes:

Horse Chestnut Extract (wt:wt) of between about 2:5 and about 8:1 , e.g., about 1 .5:1 .

[0126] 8. The composition of any of the preceding embodiments, comprising enzymes:

Peppermint Leaf Extract (wt:wt) of between about 1 :2 and about 8:1 , e.g., about 2:1 .

[0127] 9. The composition of any of the preceding embodiments, comprising enzymes: deglycyrrhizinated licorice (wt:wt) of between about 1 :1.5 and about 21 :1 , e.g., about 5.3:1.

[0128] 10. The composition of any of the preceding embodiments, comprising:

• Amylase;

• Cellulase;

• Lactase;

• Lipase;

• Protease;

• Bromelain;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin C; and

• L. plantarum.

[0129] 11. The composition of any of the preceding embodiments, comprising:

Amylase; Cellulase;

• Lactase;

• Lipase;

• Protease;

• Apple pectin;

• Bromelain;

• Butyrate;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Ginger Root Extract;

• Resveratrol;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin C; and

• L. plantarum

[0130] 12. The composition of any of the preceding embodiments for use in preparing a formulation for improving gastrointestinal health or relieving gastrointestinal discomfort.

[0131] 13. The composition of any of the preceding embodiments wherein the ingredients, together, improve at least any of at least any of one, two, three, four, five, six, seven, eight, nine, ten or eleven of the following: low digestive efficiency, high inflammatory activity, high bacterial LPS biosynthesis, increased expression of flagellar assembly pathways, low digestive efficiency pathways, high protein fermentation pathways, low intestinal barrier health pathways, high biofilm formation pathways, high prevalence of virus taxa, low microbial diversity, and low numbers of butyrate producers.

[0132] 14. A composition comprising at least any of two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or sixteen ingredients selected from:

• Amylase (e.g., about 1 ,000 DU to about 10,000 DU; about 5,000 DU to about 7,000 DU; about 6,000 DU;

• Cellulase (e.g., about 50 CU to about 1000 CU; about 25 to about 500 CU; about 275 CU;

• Lactase (e.g., about 100 ALU to about 1500 ALU; about 750 ALU to about 1250 ALU; about 1000 ALU; • Lipase (e.g., about 10 FIP to about 250 FIP; about 20 FIP to about 70 FIP; about 50 FIP;

• Protease (e.g., about 100 PC to about 2000 PC; about 1250 PC to about 1750 PC; about 1500 PC;

• Apple pectin (e.g., about 100 mg to about 20 g; about 200 mg to about 600 mg; about 400 mg;

• Bromelain (e.g., about 10 GDU to about 1000 GDU; about 10 GDU to about 100 GDU; about 50 GDU;

• Butyrate (e.g., about 50 mg to about 1500 mg; about 100 mg to about 500 mg; about 300 mg;

• Horse Chestnut Seed Extract (e.g., about 50 mg to about 1000 mg; about 50 mg to about 700 mg; about 250 mg; 470 mg;

• Peppermint Leaf Extract (e.g., about 50 mg to about 1500 mg; about 100 mg to about 700 mg; about 200 mg; 400 mg;

• Ginger Root Extract (e.g., about 10 mg to about 1500 mg; about 25 mg to about 700 mg; about 100 mg;

• Resveratrol (e.g., about 5 mg to about 1500 mg; about 25 mg to about 500 mg; about 75 mg;

• Deglycyrrhizinated Licorice (DGL) Root Extract (e.g., about 6 mg to about 1500 mg; about 26 mg to about 500 mg; about 75 mg; 150 mg;

• Vitamin A (e.g., about 0.2 mg to about 10 mg; about 0.5 mg to about 5 mg; about 2 mg;

• Vitamin C (e.g., about 20 mg to about 2000 mg; about 20 mg to about 750 mg; about 90 mg;

• Lacto aboutbacillus Postbiotic (e.g., about 5 B CFU to about 50 B CFU; about 10 B CFU to about 30 B CFU; about 20 B CFU;

• L. plantarum (e.g., about 2.5 B CFU to about 25 B CFU; about 5 B CFU to about 20 B CFU; about 10B CFU.

[0133] 15. A dose of a composition of any of embodiments 1-14, comprised in one or a plurality of dosage forms, wherein the ingredients, if present, are present in about any of the amounts set forth in Table 1 .

[0134] 16. The dose of embodiment 15, comprising ingredients of Digestive Formula

3.2, Digestive Formula 3.4, Digestive Formula 4.2, or Digestive Formula 4.4, wherein the amounts of each ingredient are the amounts or are about the amounts set forth in FIGs. 1 , 2, and 3.

[0135] 17. The dose of any of embodiments 15-16, comprised in one, two, three, four, five or six dosage forms.

[0136] 18. The dose of any of embodiments 15-16, wherein at least one of the dosage forms has a mass between about 0.5 gms and about 10 gms, e.g., between about 1 gm and 6 about gms, e.g. about 3 gms.

[0137] 19. The dose of any of embodiments 15-18, formulated for oral administration.

[0138] 20. The dose of any of embodiments 15-18, wherein all of the ingredients are included in at least one of the dosage forms.

[0139] 21 . The dose of any of embodiments 15-18, wherein a probiotic, if present, is in a different container than other ingredients.

[0140] 22. The dose of any of embodiments 15-18, wherein none of the dosage forms includes all of the ingredients.

[0141] 23. The dose of any of embodiments 15-18, wherein the dosage form or forms are selected from a capsule, a tablet, a gummy, a stick pack (e.g., pixie stick), a blister pack and a packet.

[0142] 24. The dose of any of embodiments 15-18, wherein the formulation is essentially free of milk, casein, egg, shellfish, tree nut, peanut, wheat, gluten, artificial flavor, artificial color, preservative, magnesium stearate, soybean, or an animal product.

[0143] 25. A method for improving gastrointestinal health or relieving gastrointestinal discomfort in a subject, the method comprising administering to the subject:

(a) the composition of any of embodiment 1-14, or

(b) the dose of either of embodiments 15-24.

[0144] 26. The method of embodiment 25, wherein the improvement in gastrointestinal health is alteration of gut microbial activity.

[0145] 27. The method of embodiment 25, wherein the alteration of gut microbial activity comprises increased diversity of gut microbiome.

[0146] 28. The method of embodiment 25, wherein the gastrointestinal discomfort is one or more of:

(a) abdominal pain,

(b)constipation, (c) diarrhea,

(d) cramping,

(e) bloating and

(f) gas.

[0147] 29. The method of embodiment 25, wherein the effective amount is a daily dose of each ingredient, if present in the composition, in about any of the amounts set forth in Table 1.

[0148] 30. The method of any of embodiments 25-29, comprising administering the composition daily for at least 5 days, a week or a month.

[0149] 31. The method of any of embodiments 25-29, comprising administering the composition daily for at least any of 3, 4, 5, 6 or 7 days a week for at least any of one month, two months, three months, six months or one year.

[0150] 32. The method of any of embodiments 25-29, wherein the composition is administered as a dose of any of embodiments 15-24.

[0151] 33. A kit comprising:

(a) the composition of any of embodiment 1-14, or

(b) the dose of either of embodiments 15-24.

[0152] 34. The kit of embodiment 33, further comprising instructions for use and/or dosage schedules.

EXAMPLES

Ingredient Selection Criteria

[0153] A cohort of subjects with symptoms of gastrointestinal discomfort are studied to determine which dysregulated gut microbial pathways are characteristic of these individuals, using machine learning algorithms. Ingredients are selected that positively influence these pathways and/or are known to treat symptoms of gastrointestinal discomfort.

[0154] Population statistics from the artificial intelligence recommendation engine are used to identify ingredients that were recommended to users that meet the inclusion criteria that reduced the severity of their symptom(s).

[0155] Ingredients that meet one or more of this criteria were prioritized. The ingredient list and dosages were finalized based on clinical discretion.

[0156] As used herein, the following meanings apply unless otherwise specified. The words “can” and “may” are used in a permissive sense (i.e. , meaning having the potential to), rather than the mandatory sense (i.e., meaning must). The words “include”, “including”, and “includes” and the like mean including, but not limited to. The singular forms “a,” “an,” and “the” include plural referents. Thus, for example, reference to “an element” includes a combination of two or more elements, notwithstanding use of other terms and phrases for one or more elements, such as “one or more.” The phrase “at least one” includes “one”, “one or more”, “one or a plurality”, and, therefore contemplates the use of the term “a plurality”. The term “or” is, unless indicated otherwise, non-exclusive, i.e., encompassing both “and” and “or.” The term “any of” between a modifier and a sequence means that the modifier modifies each member of the sequence. So, for example, the phrase “at least any of 1 , 2 or 3” means “at least 1 , at least 2 or at least 3”. The term “about” refers to a range that is 5% plus or minus from a stated numerical value within the context of the particular usage. The term "consisting essentially of refers to the inclusion of recited elements and other elements that do not materially affect the basic and novel characteristics of a claimed combination.

[0157] It should be understood that the description and the drawings are not intended to limit the invention to the particular form disclosed, but to the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present invention as defined by the appended claims. Further modifications and alternative embodiments of various aspects of the invention will be apparent to those skilled in the art in view of this description. Accordingly, this description and the drawings are to be construed as illustrative only and are for the purpose of teaching those skilled in the art the general manner of carrying out the invention. It is to be understood that the forms of the invention shown and described herein are to be taken as examples of embodiments. Elements and materials may be substituted for those illustrated and described herein, parts and processes may be reversed or omitted, and certain features of the invention may be utilized independently, all as would be apparent to one skilled in the art after having the benefit of this description of the invention. Changes may be made in the elements described herein without departing from the spirit and scope of the invention as described in the following claims.

[0158] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

Claims

WHAT IS CLAIMED IS:

1. A composition comprising:

(a) a Lactobacillus Postbiotic;

(b) any three, four, five or six enzymes selected from:

(i) an amylase,

(ii) a cellulase,

(iii) a lactase,

(iv) a lipase,

(v) a protease, and

(vi) bromelain; and

(c) any one, two or three extracts selected from:

(i) horse chestnut seed extract,

(ii) peppermint leaf extract, and

(iii) deglycyrrhizinated licorice.

2. The composition of claim 1, comprising all six of the enzymes of (b).

3. The composition of claim 1 or 2, comprising all three of the extracts of (c).

4. The composition of any of the preceding claims, further comprising one or both of:

(a) vitamin A, and

(b) vitamin C.

5. The composition of any of the preceding claims, further comprising one or more of:

(a) apple pectin;

(b) butyrate;

(c) ginger root extract; and

(d) resveratrol.

6. The composition of any of the preceding claims, comprising enzymes: Lactobacillus Postbiotic (gms:B CFU) of between about 5:1 and about 80:1 , e.g., about 5:1.

7. The composition of any of the preceding claims, comprising enzymes: Horse Chestnut Extract (wt:wt) of between about 2:5 and about 8:1 , e.g., about 1 .5:1.

8. The composition of any of the preceding claims, comprising enzymes: Peppermint Leaf Extract (wt:wt) of between about 1 :2 and about 8:1 , e.g., about 2:1.

9. The composition of any of the preceding claims, comprising enzymes: deglycyrrhizinated licorice (wt:wt) of between about 1 :1.5 and about 21 :1 , e.g., about 5.3:1.

10. The composition of any of the preceding claims, comprising:

• Amylase;

• Cellulase;

• Lactase;

• Lipase;

• Protease;

• Bromelain;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin C; and

• L. plantarum.

11. The composition of any of the preceding claims, comprising:

• Amylase;

• Cellulase;

• Lactase;

• Lipase;

• Protease;

• Apple pectin;

• Bromelain;

• Butyrate;

• Horse Chestnut Seed Extract;

• Peppermint Leaf Extract;

• Ginger Root Extract;

• Resveratrol;

• Deglycyrrhizinated Licorice (DGL) Root Extract;

• Vitamin C; and

• L. plantarum

12. The composition of any of the preceding claims for use in preparing a formulation for improving gastrointestinal health or relieving gastrointestinal discomfort.

13. The composition of any of the preceding claims wherein the ingredients, together, improve at least any of at least any of one, two, three, four, five, six, seven, eight, nine, ten or eleven of the following: low digestive efficiency, high inflammatory activity, high bacterial LPS biosynthesis, increased expression of flagellar assembly pathways, low digestive efficiency pathways, high protein fermentation pathways, low intestinal barrier health pathways, high biofilm formation pathways, high prevalence of virus taxa, low microbial diversity, and low numbers of butyrate producers.

14. A composition comprising at least any of two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or sixteen ingredients selected from:

• Lactase (e.g., about 100 ALU to about 1500 ALU; about 750 ALU to about 1250 ALU; about 1000 ALU;

• Lipase (e.g., about 10 FIP to about 250 FIP; about 20 FIP to about 70 FIP; about 50 FIP;

• Vitamin A (e.g., about 0.2 mg to about 10 mg; about 0.5 mg to about 5 mg; about 2 mg; • Vitamin C (e.g., about 20 mg to about 2000 mg; about 20 mg to about 750 mg; about 90 mg;

15. A dose of a composition of any of claims 1-14, comprised in one or a plurality of dosage forms, wherein the ingredients, if present, are present in about any of the amounts set forth in Table 1.

16. The dose of claim 15, comprising ingredients of Digestive Formula 3.2, Digestive Formula 3.4, Digestive Formula 4.2, or Digestive Formula 4.4, wherein the amounts of each ingredient are the amounts or are about the amounts set forth in FIGs. 1 , 2, and 3.

17. The dose of any of claims 15-16, comprised in one, two, three, four, five or six dosage forms.

18. The dose of any of claims 15-16, wherein at least one of the dosage forms has a mass between about 0.5 gms and about 10 gms, e.g., between about 1 gm and 6 about gms, e.g. about 3 gms.

19. The dose of any of claims 15-18, formulated for oral administration.

20. The dose of any of claims 15-18, wherein all of the ingredients are included in at least one of the dosage forms.

21. The dose of any of claims 15-18, wherein a probiotic, if present, is in a different container than other ingredients.

22. The dose of any of claims 15-18, wherein none of the dosage forms includes all of the ingredients.

23. The dose of any of claims 15-18, wherein the dosage form or forms are selected from a capsule, a tablet, a gummy, a stick pack (e.g., pixie stick), a blister pack and a packet.

24. The dose of any of claims 15-18, wherein the formulation is essentially free of milk, casein, egg, shellfish, tree nut, peanut, wheat, gluten, artificial flavor, artificial color, preservative, magnesium stearate, soybean, or an animal product.

25. A method for improving gastrointestinal health or relieving gastrointestinal discomfort in a subject, the method comprising administering to the subject:

(a) the composition of any of claim 1-14, or

(b) the dose of either of claims 15-24.

26. The method of claim 25, wherein the improvement in gastrointestinal health is alteration of gut microbial activity.

27. The method of claim 25, wherein the alteration of gut microbial activity comprises increased diversity of gut microbiome.

28. The method of claim 25, wherein the gastrointestinal discomfort is one or more of:

(a) abdominal pain,

(b)constipation,

(c) diarrhea,

(d) cramping,

(e) bloating and

(f) gas.

29. The method of claim 25, wherein the effective amount is a daily dose of each ingredient, if present in the composition, in about any of the amounts set forth in Table 1 .

30. The method of any of claims 25-29, comprising administering the composition daily for at least 5 days, a week or a month.

31. The method of any of claims 25-29, comprising administering the composition daily for at least any of 3, 4, 5, 6 or 7 days a week for at least any of one month, two months, three months, six months or one year.

32. The method of any of claims 25-29, wherein the composition is administered as a dose of any of claims 15-24.

33. A kit comprising:

(a) the composition of any of claim 1-14, or

(b) the dose of either of claims 15-24.

34. The kit of claim 33, further comprising instructions for use and/or dosage schedules.