Postbiotic and herbal blend for regulating metabolic processes and promoting weight loss

Abstract

The present disclosure relates to a postbiotic composition that delivers a mixture or blend of postbiotic components in combination with micronutrients, said postbiotic components derived from beneficial bacteria from Lactiplantibacillus spp. or Lactobacillus spp. (L. plantarum, L. acidophilus, and L. rhamnosus). The postbiotic compositions of the disclosure may either be a live organism capable of proliferation in the subject, a non-living strain such as, but not limited to, a heat-killed proliferative strain, or a combination thereof. In addition to the postbiotic derived from Lactiplantibacillus or Lactobacillus beneficial bacterial strains, the compositions include micronutrients such as Vitamin D, vitamin B12, and herbal extracts in various combinations to ameliorate gut and metabolic health of a human subject.

Description

POSTBTOTIC AND HERBAL BLEND FOR REGULATING METABOLIC PROCESSES AND PROMOTING WEIGHT LOSS TECHNICAL FIELD

[0001] This invention relates to postbiotic and herbal extract supplement blends for ameliorating metabolic diseases, particularly insulin resistance, pre-diabetes, and Type 2 diabetes mellitus. Postbiotics are bacteria-based products that are nonviable substances produced by bacteria or are derived from inactivated bacteria.BACKGROUND

[0002] The increasing incidence and prevalence of obesity and metabolic diseases is a significant public health concern, necessitating ongoing research into effective therapies and preventative strategies. The gut microbiome plays a critical role in regulating metabolic processes and can be modulated to support healthy metabolism and a healthy weight.

[0003] The next generation of bacteria-based products, known as postbiotics, are nonviable substances produced by bacteria or derived from inactivated bacteria. Postbiotics such as heat-killed probiotic strains can have potent anti-inflammatory effects via their cell wall components and metabolites. Examples of postbiotics include heat-killed Lactobacilli, vitamin B12, vitamin K, folate, lipopolysaccharides, enzymes, and short chain fatty acids (SCFAs), representing a subset of essential nutrients. Postbiotics have been observed to demonstrate anti-obesity and anti-diabetic effects through a variety of mechanisms. These pathways primarily involve an elevation in energy expenditure, a decrease in the formation and differentiation of adipocytes, decrease in food intake, modification of lipid and carbohydrate absorption and metabolism, and regulation of gut dysbiosis. Based on these effects and mechanisms, the use of probiotics can be considered as a potential strategy for the prevention and treatment of metabolic diseases such as obesity, type 2 diabetes mellitus (T2DM), and concomitant cardiovascular complications.Postbiotics can also modulate the release of gut hormones, such as glucagon-like petide-1 (GLP-1) which is key in insulin homeostasis, glucose regulation, and triggering of feelings of satiety, critical in food intake modulation.

[0004] Micronutrient and herbal supplementation have been historically harnessed for metabolic support. Chromium picolinate (CrP) has been used to treat chromium deficiency, as an aid to controlling blood sugar in people with diabetes and prediabetes, to lower cholesterol, and as a weight-loss supplement. Randomized controlled clinical research suggest that CrP consumptionmay result in the reduction of body weight. Consumption of white mulberry extract or mulberry leaf-based foods modulate the gut microbiome and affect expression of genes related to obesity. Extensive preclinical and clinical research have outlined the potential uses of fenugreek for its anti-diabetic, anti-hyperlipidemic, anti-obesity, anti-cancer, anti-inflammatory, antioxidant, antifungal, and anti-bacterial effects. Additional micronutrients such as vitamins can support gut health directly while also having a distal effect on organs such as the heart, brain, and lungs.

[0005] The rapidly rising incidence of obesity, coupled with T2DM, is a growing public health concern. Glucagon-like peptide 1 (GLP-1), an endogenous peptide secreted by enteroendocrine L-cells, demonstrates exceptional pharmacological potential for the treatment of T2DM and obesity, primarily through its pivotal roles in regulating glucose homeostasis, stimulating glucose-dependent insulin secretion, and promoting satiety.

[0006] Certain nutrients are particularly useful for the prevention and treatment of metabolic diseases such as obesity, type 2 diabetes mellitus (T2DM), and concomitant cardiovascular complications. Postbiotics can modulate the release of gut hormones, such as GLP-1, which is key in insulin homeostasis, glucose regulation, and triggering of feelings of satiety, critical in food intake modulation. There is an ongoing need for compositions and formulations to treat these conditions of public health concern.

[0007] Thus, there exists a need for compositions and formulations containing postbiotics to meet these and other needs. The compositions and formulations of the present invention meet these and other needs as detailed below.SUMMARY

[0008] In one embodiment a dietary supplement, food product, or nutraceutical is described for use in supporting health in a human subject, comprising a blend of postbiotics, chromium picolinate, white mulberry extract, fenugreek extract, and vitamins B 12 and D3. The supplements described herein may be formulated for oral administration.

[0009] In one embodiment a dietary supplement, food product, or nutraceutical is described for use in supporting health in a human subject, comprising a blend of postbiotics, chromium picolinate, white mulberry extract, fenugreek extract, and vitamins B12 and D3, where the blend of postbiotics comprises RSB11® postbiotic (ResBiotic Nutrition, Inc., Birmingham, Alabama). The supplements described herein may be formulated for oral administration.

[0010] In one embodiment a dietary supplement, food product, or nutraceutical is described for use in supporting health in a human subject, comprising a blend of postbiotics, chromium picolinate, white mulberry extract, fenugreek extract, and vitamin B 12, where the blend of postbiotics comprises RSB 11® postbiotic. The supplements described herein may be formulated for oral administration.

[0011] In one embodiment a dietary supplement, food product, or nutraceutical is described for use in supporting health in a human subject, comprising a blend of postbiotics, chromium picolinate, white mulberry extract, fenugreek extract, and vitamin B 12, where the blend comprises RSB11® and resM blends. The supplements described herein may be formulated for oral administration.

[0012] In one embodiment a dietary supplement method is described for improving gastrointestinal and metabolic health comprising administering the nutritional postbiotic blend of the present invention to a human subject by oral administration.BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Further aspects of the present disclosure will be more readily appreciated upon review of the detailed description of its various embodiments, described below, when taken in conjunction with the accompanying drawings. Statistical analysis was performed using one-way ANOVA (GraphPad Prism vlO.1.2) with Dunnett’s post-hoc test. * p < 0.05; ** p < 0.01; *** p < 0.001; ****p< 0.0001.

[0014] FIG. 1A depicts a three-fold increase in GLP-1 protein (pg / mL) in Caco-2 intestinal epithelial cells after 24h incubation time as measured by GLP-1 ELISA kit in cell culture supernatant upon dosing with resM micronutrient blend. **** p < 0.0001

[0015] FIG. IB depicts glucose consumption (mg / dL) upon dosing with resM micronutrient blend. *** p < 0.001.

[0016] FIG. 2 depicts increase in GLP-1 protein (pg / mL) in Caco-2 intestinal epithelial cells after 24h incubation time as measured by GLP-1 ELISA kit in cell culture supernatant upon dosing with resM™, a blend of RSB 1 l®-heat inactivated (HI) postbiotic and the resM micronutrient blend. * p < 0.05; **** p < 0.0001.

[0017] FIG. 3A depicts increases in active GLP-1 secretion in an ex vivo model using human ileum organoids treated with resM micronutrient blend portion alone per Tables 1 & 2 (10 mg). * p < 0.05.

[0018] FTG. 3B depicts increases in active GLP-1 secretion in an ex vivo model using human ileum organoids treated with RSB11®-heat inactivated (HI) postbiotic alone per Tables 1 & 2 (3 mg). ** p < 0.01.DETAILED DESCRIPTION

[0019] The present disclosure encompasses embodiments of a postbiotic composition comprising heat killed or deactivated forms of probiotic bacteria. In one embodiment the heat killed, heat-inactivated or deactivated probiotic bacteria is Lactiplantibacillus plantarum, e.g. RSB11® HI (L. plantarum), formerly known as Lactobacillus plantarum.

[0020] Preclinical studies show that Cell Free Supernatant from Lactobacilli rhamnosus Y37, Lactobacilli plantarum XI, and L. plantarum CCFM36 decreased fasting blood glucose (FBG) and increased GLP-1 levels in a high-fat diet-induced T2DM mouse model (Li X., Xu Q., Jiang T., Fang S., Wang G., Zhao J., et al. A comparative study of the antidiabetic effects exerted by live and dead multi-strain probiotics in the type 2 diabetes model of mice. Food Funct.2016;7(12):4851-60; Mishra N., Garg A., Ashique S., and Bhatt S., Potential of postbiotics for the treatment of metabolic disorders. Drug Discov Today. 2024;29(4):10392). In another embodiment the postbiotic compositions of the invention comprise micronutrients. These micronutrients include nutritionally active ingredients.

[0021] In another embodiment, the postbiotic compositions comprise postbiotic metabolites such as short-chain fatty acids (SCFAs) that can indirectly influence appetite regulation, as chronic inflammation can disrupt normal appetite signaling (Arpaia N., Campbell C., Fan X., Dikiy S., van der Veeken J., deRoos P., et al., Metabolites produced by commensal bacteria promote peripheral regulatory T-cell generation. Nature. 2013;504(7480):451-5; Iraporda C., Errea A., Romanin D. E., Cayet D., PereyraE., Pignataro O., etal., Lactate and short chain fatty acids produced by microbial fermentation downregulate proinflammatory responses in intestinal epithelial cells and myeloid cells. Immunobiology. 2015;220(10):1161-9; ParkM., Joung M., Park J. H., Ha S. K., Park H. Y., Role of Postbiotics in Diet-Induced Metabolic Disorders. Nutrients.2022; 14(18)). Moreover, postbiotics might impact serotonin production in the gut, affecting mood and satiety signals (Cerdo T., Garcia-Santos J. A., M G. B., Campoy C., The Role of Probiotics and Prebiotics in the Prevention and Treatment of Obesity. Nutrients. 2019;11(3)). They can also modulate the release of gut hormones, such as peptide YY (PYY) and glucagon-like peptide- 1 (GLP-1), contributing to feelings of fullness and satiety (Li X., et al., supra). Furthermore,postbiotics support gut barrier function, and a healthy gut barrier is associated with improved metabolic health and energy balance (Mishra N. et al, supra).

[0022] In another embodiment, the postbiotic compositions comprise micronutrients such as Chromium picolinate (CrP) which is used as an oral treatment to help reduce body weight and alter body composition. It is an organic compound of trivalent chromium, an essential trace mineral and cofactor to insulin, and picolinic acid, a naturally occurring derivative of tryptophan. Chromium enhances insulin activity and has been the subject of several studies assessing its effects in carbohydrate, protein and lipid metabolism (Anderson R. A., Effects of chromium on body composition and weight loss. Nutr Rev. 1998;56(9):266-70; Pittier M. H., Stevinson C., and Ernst E., Chromium picolinate for reducing body weight: meta-analysis of randomized trials. Int J Obes Relat Metab Disord. 2003;27(4):522-9; Willoughby D., Hewlings S., and Kalman D., Body Composition Changes in Weight Loss: Strategies and Supplementation for Maintaining Lean Body Mass, a Brief Review. Nutrients. 2018; 10(12)). Effects include an increase in lean body mass, a decrease in percentage body fat and an increase in basal metabolic rate. In a randomized, doubleblinded, placebo-controlled 10-month study on chromium picolinate and type 2 diabetes found that supplementation with chromium picolinate significantly attenuated weight gain in subjects who were taking sulfonylurea agents (Martin J., Wang Z. Q., Zhang X. H., Wachtel D., Volaufova J., Matthews D. E., et al., Chromium picolinate supplementation attenuates body weight gain and increases insulin sensitivity in subjects with type 2 diabetes. Diabetes Care. 2006;29(8): 1826-32). Additionally, chromium picolinate might help reduce appetite and cravings, particularly in individuals with binge-eating disorders or depression (Brownley K. A., Von Hoile A., Hamer R. M., La Via M., and Bulik C. M., A double-blind, randomized pilot trial of chromium picolinate for binge eating disorder: results of the Binge Eating and Chromium (BEACh) study. J Psychosom Res. 2013;75(l):36-42).

[0023] In another embodiment, the postbiotic compositions comprise micronutrients such as white mulberry extract, particularly from the leaves of Morns alba, which has shown potential as a tool to treat obesity. White mulberry extract contains several compounds that contribute to weight loss, primarily polyphenols, dietary fiber, and certain alkaloids. Studies indicate that it can aid weight loss by inhibiting enzymes like alpha-glucosidase, which reduces carbohydrate absorption and lowers the glycemic index (Hansawasdi C., and Kawabata J., Alpha-glucosidase inhibitory effect of mulberry (Morns alba) leaves on Caco-2. Fitoterapia. 2006;77(7-8):568-73).Additionally, mulberry leaf polyphenols have been found to inhibit weight gain in high-fat diet-induced obese mice by altering lipid metabolism and increasing the expression of thermogenesis-related genes ( Li R., Zhu Q., Wang X., and Wang H., Mulberry leaf polyphenols alleviated high-fat diet-induced obesity in mice. Front Nutr. 2022;9:979058). Preclinical studies suggest that mulberry extract can modulate GLP-1 levels, glucose levels (Cheng J-J., Wei W-C., Chen C-C., Shiao Y-J., Huang N-K., Liao C-H., etal., Mulberry leaf extract exhibits multiple antidiabetic activities and alleviates dysglycemia, systemic inflammation, hepatic steatosis, and xenobiotic metabolism abnormalities in type 2 diabetic mice. Journal of Functional Foods.2024; 119: 106330). Additionally, mulberry leaf fiber and polyphenols have a synergistic effect, influencing gut microbiota and metabolites (Zheng XX., Li DX., Li YT., Chen YL., Zhao YL., Ji S., et al., Mulberry leaf water extract alleviates type 2 diabetes in mice via modulating gut microbiota-host co-metabolism of branched-chain amino acid. Phytother Res. 2023;37(8):3195-210). Specifically, mulberry leaf fiber has a synergistic effect that modulates the intestinal microbiome, restoring balance of Firmicutes to Bacteroidota in the gut (Rodriguez-Sojo M. J., Ruiz-Malagon A. J., Hidalgo-Garcia L., Molina-Tijeras J. A., Diez-Echave P., Lopez-Escanez L., et al., The Prebiotic Effects of an Extract with Antioxidant Properties from Morus alba L.Contribute to Ameliorate High-Fat Diet-Induced Obesity in Mice. Antioxidants (Basel).2023;12(4)).

[0024] In another embodiment, the postbiotic compositions comprise micronutrients such as herbal extracts from Fenugreek (Trigonella foenum-graecum) nutritional supplementation with which has been shown to positively affect gut microbiota by reversing the dysbiotic effects of high-fat diets. Animal studies indicate that fenugreek can improve glucose tolerance and lipid profiles in high-fat diet-fed mice by reducing body weight gain and plasma triglycerides (Bruce-Keller A. J., Richard A. J., Fernandez-Kim S-O., Ribnicky D. M., Salbaum J. M., Newman S., etal., Fenugreek Counters the Effects of High Fat Diet on Gut Microbiota in Mice: Links to Metabolic Benefit. Scientific Reports. 2020; 10(1): 1245). Studies indicate that fenugreek increases overall microbiome diversity. Clinical trials suggest a hypoglycemic role for fenugreek in managing T2DM (Shabil M., Bushi G., Bodige P. K., Maradi P. S., Patra B. P., Padhi B. K., et al., Effect of Fenugreek on Hyperglycemia: A Systematic Review and Meta-Analysis. Medicina (Kaunas).2023;59(2); Hassani S. S., Fallahi Arezodar F, Esmaeili S. S., and Gholami-Fesharaki M., Effect of Fenugreek Use on Fasting Blood Glucose, Glycosylated Hemoglobin, Body Mass Index, WaistCircumference, Blood Pressure and Quality of Life in Patients with Type 2 Diabetes Mellitus: A Randomized, Double-Blinded, Placebo-Controlled Clinical Trials. Galen Med J. 2019;8:el432). The phytochemical analysis of fenugreek has revealed the presence of various categories of secondary metabolites, such as saponins, steroids / diosgenin, alkaloids, flavonoids, terpenes, phenolic acid derivatives, amino acids and fatty acids and their derivatives (Shabil et al., supra; Kiss R., Pesti-Asboth G., Szarvas M. M., Stiindl L., Cziaky Z., Hegedus C., et al. Diosgenin and Its Fenugreek Based Biological Matrix Affect Insulin Resistance and Anabolic Hormones in a Rat Based Insulin Resistance Model. Biomed Res Int. 2019;2019: 7213913). Fenugreek is one of very few natural sources of diosgenin which has a wide range of anti-inflammatory and anticancer activities (Luo W., Deng J., He J., Yin L., You R., Zhang L, et al., Integration of molecular docking, molecular dynamics and network pharmacology to explore the multi-target pharmacology of fenugreek against diabetes. J Cell Mol Med. 2023;27(14): 1959-74).

[0025] In another embodiment, the postbiotic compositions comprise micronutrients such as vitamins and minerals to supplement the health benefits provided by postbiotics and herbal extracts. Vitamin B12 (in the form of methyl cobalamin) is a water-soluble vitamin that supports healthy central nervous system development and function, red blood cell production, and DNA synthesis. A double-blinded, placebo-controlled clinical trial suggests that Vitamin B12 may be useful for managing pain, improving quality of life, and improving neurological scores in patients with diabetic neuropathy (Didangelos T., Karlafti E., Kotzakioulafi E., Margariti E., Giannoulaki P., Batanis G., et al., Vitamin B12 Supplementation in Diabetic Neuropathy: A 1-Year, Randomized, Double-Blind, Placebo-Controlled Trial. Nutrients. 2021;13(2)) and in people with obesity undergoing gastric bypass (Ramos R. J., Mottin C. C., Alves L. B., Mulazzani C. M., and Padoin A. V., Vitamin B12 supplementation orally and intramuscularly in people with obesity undergoing gastric bypass. Obes Res Clin Pract. 2021;15(2):177-9).

[0026] In another embodiment, the postbiotic compositions comprise micronutrients such as vitamins B and E supplementation with which can be used to inhibit reactive oxygen species (ROS) production. Many probiotics can synthesize folates and B-complex vitamins. Dysbiosis of gut health causes excessive production of reactive oxygen species (ROS), which leads to inflammation, cellular dysfunction, and diabetes, among others. In T2DM, deficiency of folate and vitamin B12 leads to oxidative stress (van de Lagemaat E. E., de Groot L., and van den Heuvel E. Vitamin B(12) in Relation to Oxidative Stress: A Systematic Review. Nutrients. 2019;11(2)).Additionally, vitamin Bl 2 and glutathione synthetase (GSH) deficiency are linked to various oxidative stress-involved conditions, including T2DM (Abboud M., Rizk R., AlAnouti F., Papandreou D., Haidar S., and Mahboub N., The Health Effects of Vitamin D and Probiotic CoSupplementation: A Systematic Review of Randomized Controlled Trials. Nutrients. 2020; 13(1)).

[0027] In another embodiment, the postbiotic compositions comprise micronutrients such as vitamin D3, which has emerged as a focal point in enhancing insulin sensitivity (Krajewska M., Witkowska-S^dek E., RuminskaM., Stelmaszczyk-Emmel A., Sobol M., Majcher A., et al., Vitamin D Effects on Selected Anti-Inflammatory and Pro-Inflammatory Markers of Obesity-Related Chronic Inflammation. Front Endocrinol (Lausanne). 2022; 13:920340), modulating lipid profiles (Mei L. H., Zheng W. X., Zhao Z. T., Meng N., Zhang Q. R., Zhu W. J., et al., A Pilot Study of the Effect of Lactobacillus casei Obtained from Long-Lived Elderly on Blood Biochemical, Oxidative, and Inflammatory Markers, and on Gut Microbiota in Young Volunteers. Nutrients. 2021; 13(11)), and exerting anti-inflammatory effects (Milajerdi A., Mousavi S. M., Sadeghi A., Salari-Moghaddam A., Parohan M., Larijani B., etal., The effect of probiotics on inflammatory biomarkers: a meta-analysis of randomized clinical trials. Eur J Nutr.2020;59(2):633-49), while probiotics have been recognized for their ability to modulate the gut microbiota (Cho Y. A., Kim J., Effect of Probiotics on Blood Lipid Concentrations: A MetaAnalysis of Randomized Controlled Trials. Medicine (Baltimore). 2015;94(43):el714), systemic inflammation (Mason C., Xiao L., Imayama L, Duggan C., Wang C. Y., Korde L., et al., Vitamin D3 supplementation during weight loss: a double-blind randomized controlled trial. Am J Clin Nutr. 2014;99(5): 1015-25), and lipid metabolism (Xenos K., Papasavva M., Raptis A., Katsarou M. S., and Drakoulis N., Vitamin D Supplementation and Genetic Polymorphisms Impact on Weight Loss Diet Outcomes in Caucasians: A Randomized Double-Blind Placebo-Controlled Clinical Study. Front Med (Lausanne). 2022;9:811326). A recent systematic review of seven clinical randomized controlled trials (RCTs) underscored the synergistic benefits of vitamin D and probiotic co-supplementation (lactic acid bacteria) in ameliorating chronic disease markers and improving metabolic health more effectively than either component alone (Musazadeh V., Kavyani Z., Mirhosseini N., Dehghan P., and Vajdi M., Effect of vitamin D supplementation on type 2 diabetes biomarkers: an umbrella of interventional meta-analyses. Diabetol Metab Syndr.2023; 15(1):7). L. casei and L. rhamnosus and Vitamin D3 enriched Orange Juice led to weight loss, less energy / macronutrient consumption, improved lipid profiles, and increased insulinsensitivity after 8 weeks in those following a Westernized diet (Papakonstantinou E., Zacharodimos N., Georgiopoulos G., Athanasaki C., Bothou D. L., Tsitsou S., et al., Two-Month Consumption of Orange Juice Enriched with Vitamin D3 and Probiotics Decreases Body Weight, Insulin Resistance, Blood Lipids, and Arterial Blood Pressure in High-Cardiometabolic-Risk Patients on a Westernized Type Diet: Results from a Randomized Clinical Trial. Nutrients.2024; 16(9)).

[0028] In another embodiment, the present disclosure encompasses embodiments of a postbiotic composition that delivers postbiotics preferably derived from Lactiplantibacillus plantarum (RSB11®). In another embodiment, the postbiotic compositions are derived from beneficial bacteria Lactobacillus acidophilus (RSB12®), or Lactobacillus rhamnosus (RSB13®). The postbiotic compositions of the disclosure comprise these beneficial bacterial strains that may either be a live organism capable of proliferation in the subject, a non-living strain such as, but not limited to, a heat-killed proliferative strain, or a combination thereof. In addition to the Lactiplantibacillus or Lactobacillus strains, the compositions can include micronutrients such as Vitamins B12 and D. These micronutrients include nutritionally active ingredients.

[0029] In another embodiment, the postbiotics are derived from any one of or any combination of the seven bacterial strains cited above and may be included in combination as a blend, for example, as resM micronutrient blend comprising chromium picolinate, white mulberry extract, fenugreek extract, vitamin Bl 2, and vitamin D3.

[0030] In another embodiment, the postbiotics are derived from heat-inactivated Lactiplantibacillus plantarum (RSB11®HI) and may be included in combination as a blend, for example, as resM blend comprising chromium picolinate, white mulberry extract, fenugreek extract, vitamin Bl 2, and vitamin D3.

[0031] In another embodiment, the postbiotics derived from any one of or any combination of the seven bacterial strains cited above may be included in combination as a blend, for example, as 35-40 mg of heat-inactivated Lactiplantibacillus plantarum (RSB11®HI) postbiotic (1.0-2.0×1010cells), 460 mg of resM blend comprising chromium picolinate 600 mcg, white mulberry extract 250 mg, fenugreek extract 200 mg, vitamin B 12200 mcg, and vitamin D3 10 mcg / 400 IU.

[0032] Each of the above postbiotic compositions, such as RSB11®, Lactiplantibacillus plantarum (RSB11®HI), and resM blend are available from ResBiotic Nutrition, Inc. (Birmingham, Alabama).

[0033] RSB11®HI is a next-generation postbiotic that delivers live-equivalent or superior efficacy in a non-viable, stable composition, retaining and enhancing the immunomodulatory power of live probiotics while eliminating the limitations of viability. This establishes RSB11-HI as a safe, stable, and efficacious postbiotic platform with broad anti-inflammatory potential. For example, the form is generally in a freeze-dried, heat-inactivated bacterial powder, having an effective concentration of about 3 mg / mL (across epithelial and non-epithelial models). When tested in a challenge model using LPS (5 pg / mL) or E. coli (107CFU / mL) for 4 h and measuring for the following endpoints: qPCR (MMP-9), ELISA (TNF-a, IL-6, IL-ip), this postbiotic demonstrated superior anti-inflammatory characteristics relative to its live counterpart. In human cell line tests (Caco-2, HBE, BG1, MG63, A498, HepG2) RSB11-HI demonstrated equivalent or superior efficacy in protease suppression, or inflammatory cytokine reduction, compared to its live counterpart, confirming that heat inactivation preserves and may enhance functional bioactivity.

[0034] For example, RSB11®HI significantly suppressed MMP-9 expression across multiple tissue models (bone, liver, gut, lung, ovary) under inflammatory stress (LPS / E. coli challenge), indicating improved metabolic balance and reduced catabolic remodeling.

[0035] In another embodiment, the postbiotic compositions can be administered with probiotics or bacterial extracts via oral administration.

[0036] In another embodiment, a postbiotic “bacterial extract” is derived from Lactiplantibacillus plantarum (RSB11® or RSB11®HI). In another embodiment, the postbiotic compositions are derived from beneficial bacteria Lactobacillus acidophilus (RSB12®), and Lactobacillus rhamnosus (RSB13®).

[0037] Other beneficial bacterial strains include, but are not limited to, L. plantarum Lp-202195, L. acidophilus NCFM, L. rhamnosus GG, and L. rhamnosus HN001. Any of the aforementioned strains may be used in the blends described herein.

[0038] The Lactobacillus genus is extremely diverse and expanding every year. With over 230 species, it has grown into one of the biggest genera in the bacterial taxonomy. As the genus has exceeded the acceptable “normal diversity,” renaming and re-classification is inevitable wherein the genus Lactobacillus may be split into most likely twelve new genera. For example, in the present disclosure, the new genus Lactiplantibacillus has been designated to replace at least one postbiotic heated-inactivated strain, formerly designated as a Lactobacillus strain. Many traditional “probiotic” species with substantiated industrial importance and starter cultures manyno longer eventually be called “ actobacillus ” Hence, a substantial communication challenge looms ahead to reduce the inevitable confusion regarding the “old commercial” and “correct scientific” nomenclature. Once the International Committee on Systematics of Prokaryotes publishes new nomenclature in their official journal, the INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, the changes are valid and official. The manuscript that will be submitted for publication outlining the new nomenclature of the Lactobacillus genus will likely be ready for submission by the end of 2018. Meanwhile, there was a taxonomic subcommittee meeting in September 2018 to discuss the nomenclature changes and an (invite-only) expert LAB IP workshop in October 2018 that evaluated the science while considering the consequences for regulations, legal / IP, and industry.

[0039] Probiotics are measured by colony forming units (“CFUs”). Few studies have been done to determine effective dosages, but effective dosages are usually in the hundreds of millions of CFUs or higher. If probiotics are being used to help with digestion, probiotics should be taken with meals, but otherwise the probiotics may survive better if taken between meals, particularly if taken with liquids that help to dilute stomach acid and move the probiotics more quickly into the digestive tract. Probiotics may be given short-term (e.g., in a daily dose for days or weeks) or long-term (over several months, or more). In the case of postbiotics, which are bacteria-based products that are nonviable substances produced by bacteria or are derived from inactivated bacteria, CFU counts are taken to quantify the number of live cells prior to inactivation of the cells by heat or other means to produce the postbiotic. A postbiotic’s concentration is measured by cells / g, or from the total number of cells used to produce it i.e. billions (109) of cells.

[0040] In some implementations of postbiotic compositions, the concentration of the probiotic microorganism (measured in CFU / g) prior to generating the postbiotics (measured in cells / g) in the composition may be at least about 1·109CFU / g, at least about 2·109CFU / g, at least about 3·109CFU / g, at least about 4·109CFU / g, at least about 5·109CFU / g, at least about 6·109CFU / g, at least about 7·109CFU / g, at least about 8·109CFU / g, at least about 9·109CFU / g, at least about 1·1010CFU / g, at least about 1.1·1010CFU / g, at least about 1.2·1010CFU / g, at least about 1.3·1010CFU / g, at least about 1.4·1010CFU / g, at least about 1.5·1010CFU / g, at least about 1.6·1010CFU / g, at least about 1.7·1010CFU / g, at least about 1.8·1010CFU / g, at least about 1.9·1010CFU / g, at least about 2·1010CFU / g, at least about 3·1010CFU / g, at least about 4·1010CFU / g, at least about 5·1010CFU / g, at least about 6·1010CFU / g, at least about 7·1010CFU / g, atleast about 8·1010CFU / g, at least about 9·1010CFU / g, or at least about 1·1011CFU / g, or at least about 2·1011CFU / g, at least about 3·1011CFU / g, at least about 4·1011CFU / g, at least about 5·1011CFU / g, at least about 6·1011CFU / g, at least about 7·1011CFU / g, at least about 8·1011CFU / g, at least about 9·1011CFU / g, or at least about 1·1012CFU / g.

[0041] As used herein, an “effective amount” or an “amount effective for” is defined as an amount effective, at dosages and for periods of time necessary, to achieve a desired biological result, such as reducing, preventing, or treating a disease or condition and / or inducing a particular beneficial effect. The effective amount of compositions of the disclosure may vary according to factors such as age, sex, and weight of the individual. Dosage regime may be adjusted to provide the optimum response. Several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of an individual’s situation. As will be readily appreciated, a composition in accordance with the present disclosure may be administered in a single serving or in multiple servings spaced throughout the day. As will be understood by those skilled in the art, servings need not be limited to daily administration, and may be on an every second or third day or other convenient effective basis. The administration on a given day may be in a single serving or in multiple servings spaced throughout the day depending on the exigencies of the situation.

[0042] The postbiotic and micronutrient blend for gut health of the present invention is unique in that it combines per unit dose a spectrum of postbiotic components comprising Lactiplantibacillus plantarum RSB11® HI postbiotic, and micronutrient and herbal extract blend resM micronutrient blend comprising micronutrients chromium picolinate, vitamin B12, and vitamin D3, and herbal extracts white mulberry extract, and fenugreek extract. Administration methods may be orally dosed directly, mixed in food or drink as dry powder or liquid, or delivered in a solid or chewable forms. The compositions of the present invention serve to seed the intestinal microbiome with beneficial bacteria and micronutrients known to improve gut microbiomes and useful for regulating metabolic processes and modulate the microbiome to support a healthy weight of a human subject.

[0043] The postbiotic and micronutrient blend for gut health dietary supplement of the present invention comprises per unit dose postbiotics derived from Lactiplantibacillus plantarum RSB11® HI present in up to about 10 billion (B) cells (or, up to 1 IB, or 12B, or 13B, or 14B, or 15B, or 16B, or 17B, or 18B, or 19B, or 20B), a micronutrient and herbal blend (resM blend)present in an amount up to about 1,000 mg (up to 2,000mg, or 3,000 mg, or 4,000 mg, or 5,000 mg), comprising micronutrients chromium picolinate present in an amount up to about 1000 mcg, vitamin B 12 is present in an amount up to about 200 mcg to 400 mcg, and vitamin D3 is present in an amount up to about 20 mcg or 800 IU, and herbal extracts comprising white mulberry extract is present in an amount up to about 500 mg, and the fenugreek extract is present in an amount up to about 500 mg.

[0044] The method described herein effects maintenance of healthy gut microflora in an individual, such as a human subject, by promoting production of gut hormones to regulate glucose homeostasis, stimulating glucose dependent insulin secretion, and promoting satiety.

[0045] In certain embodiments, the compositions comprising one or more postbiotics can include one or more dry carriers selected from the group consisting of trehalose, maltodextrin, rice flour, microcrystalline cellulose, magnesium stearate, inositol, fructooligosaccharide, galactooligosaccharide, dextrose, and the like. In certain embodiments, the dry carrier can be added to the compositions comprising one or more of the above bacterial strains in a weight percentage of from about 1% to about 95% by weight of the composition.

[0046] In certain embodiments, the compositions comprising one or more of the above postbiotic components can include one or more liquid or gel-based carriers, selected from the group consisting of water and physiological salt solutions, urea, alcohols and derivatives thereof (e.g., methanol, ethanol, propanol, butanol), glycols (e.g., ethylene glycol, propylene glycol), and the like; natural or synthetic flavorings and food-quality coloring agents, all compatible with the organism; thickening agents selected from the group consisting of corn starch, guar gum, xanthan gum, and the like; one or more spore germination inhibitors selected from the group consisting of hyper-saline carriers, methylparaben, guar gum, polysorbate, preservatives, and the like. In certain embodiments, the one or more liquid or gel-based carrier(s) can be added to the compositions comprising one or more of the above bacterial strains in a weight / volume percentage of from about 0.6% to about 95% weight / volume of the composition. In certain embodiments, the natural or synthetic flavoring(s) can be added to the compositions comprising one or more of the above bacterial strains in a weight / volume percentage of from about 3.0% to about 10.0% weight / volume of the composition. In certain embodiments, the coloring agent(s) can be added to the compositions comprising one or more of the above bacterial strains in a weight / volume percentage of from about 1.0% to about 10.0% weight / volume of the composition. In certain embodiments,the thickening agent(s) can be added to the compositions comprising one or more of the above bacterial strains in a weight / volume percentage of about 2% weight / volume of the composition.

[0047] Delivery Systems

[0048] Suitable dosage forms include tablets, capsules, solutions, suspensions, powders, gums, and confectionaries. Sublingual delivery systems include, but are not limited to, dissolvable tabs under and on the tongue, liquid drops, and beverages. Edible films, hydrophilic polymers, oral dissolvable films, or oral dissolvable strips can be used.

[0049] For oral administration, postbiotics may be further combined with one or more solid inactive ingredients for the preparation of tablets, capsules, pills, powders, granules, or other suitable dosage forms. For example, the active agent may be combined with at least one excipient selected from the group consisting of fillers, binders, humectants, disintegrating agents, solution retarders, absorption accelerators, wetting agents, absorbents, and lubricating agents. Other useful excipients include, but are not limited to, magnesium stearate, calcium stearate, mannitol, xylitol, sweeteners, starch, carboxymethylcellulose, microcrystalline cellulose, silica, gelatin, silicon dioxide, and the like.

[0050] The components of the compositions administered according to the methods of the present disclosure can be administered in a wide variety of oral dosage forms. It will be obvious to those skilled in the art that the following dosage forms may comprise, in certain embodiments, as the active component, either a chemical compound of the present disclosure or an acceptable salt of a chemical compound of the present disclosure.

[0051] For preparing nutraceutical compositions to be administered according to the methods of the present disclosure, nutraceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, and cachets. A solid carrier can be one or more substances that may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or encapsulating materials.

[0052] In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired.

[0053] In certain embodiments, powders and tablets administered according to methods of the present disclosure preferably may contain from five or ten to about seventy percent of the active compounds. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar,lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term “preparation” is intended to include the formulation of the active compound with encapsulating material as carrier providing a capsule in which the active component, with or without additional carriers, is surrounded by a carrier, which is thus in association with it. Similarly, tablets, powders, capsules, pills, sachets, and lozenges are included. Tablets, powders, capsules, pills, sachets, and lozenges can be used as solid forms suitable for oral administration.

[0054] Liquid preparations include, but are not limited to, solutions, suspensions, and emulsions, for example, water or water-propylene glycol solutions. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulation agents such as suspending, stabilizing, and / or dispersing agents. Oil carriers include, but are not limited to, sunflower oil, cranberry seed oil, algal oil, palm oil, coconut oil, and rice bran oil. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

[0055] Aqueous solutions suitable for oral use can be prepared by dissolving the active components in water and adding suitable colorants, flavors, stabilizing and thickening agents, as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0056] Compositions suitable for topical administration in the mouth, or buccal, or sublingual administration include, but are not limited to: lozenges comprising the active agent in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in suitable liquid carrier.

[0057] The nutraceutical preparations are preferably in unit dosage forms. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active components. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packaged tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, sachet, or lozenge itself; or it can be the appropriate number of any of these in packaged form.

[0058] Tablets, capsules, and lozenges for oral administration and liquids for oral use are preferred dosage form compositions.

[0059] Further details on techniques for formulation and administration may be found in the latest edition of REMINGTON’ S PHARMACEUTICAL SCIENCES (Mack Publishing Co., Easton, PA).

[0060] Routes of Administration

[0061] The compositions and compounds may be administered by any route, including, but not limited to, oral, sublingual, buccal, or as an oral spray.

[0062] The methods described above may be further understood in connection with the following Examples. In addition, the following non-limiting examples are provided to illustrate the invention. However, the person skilled in the art will appreciate that it may be necessary to vary the procedures for any given embodiment of the invention, e.g, vary the order or steps.

[0063] In one embodiment, the addition of micronutrients chromium picolinate, Vitamin B12 and vitamin D3 and herbal extracts of white mulberry extract and fenugreek extract to a blend of postbiotic compositions derived from Lactobacillus spp. or Lactiplantibacillus spp. of the invention are useful as a preventative supplement to ameliorate the incidence of obesity and metabolic diseases. The following nonlimiting combinations of strains and nutrients are contemplated herein.

[0064] The invention described herein is embodied in the following non-limiting examples. In the examples below, the term “B cells” is defined as billion(s) (109). of cells.EXAMPLE 1

[0065] resM Formulation (first variant)

[0066] Table 1 shows one preferred combination of a postbiotic composition as described herein. The postbiotic resM composition comprises Lactiplantibacillus plantarum (RSB11® HI) cells heat killed at 100 ° C plus the resM blend micronutrient composition. The resM micronutrient blend composition of the invention comprises chromium picolinate, white mulberry extract, fenugreek extract, vitamin Bl 2, and vitamin D3 as detailed in Table 1 below.TABLE 11Source: Indovedic Nutraceuticals Pvt. Ltd., Bangalore, India; starting material: Mulberry leaves (Morus alba), country of origin: India, standardized to 2% 1-deoxynojirimycin (1-DNJ).1'1' Source: Indovedic Nutraceuticals Pvt. Ltd., Bangalore, India; starting material: Fenugreek seeds (Trigonella foenum-graecum), country of origin: India, standardized to 25% total saponins (dried basis).EXAMPLE 2

[0067] resM Formulation (second variant)

[0068] Table 2 shows another preferred combination of a postbiotic composition as described herein. The postbiotic resM composition comprises Lactiplantibacillus plantarum (RSB11® HI) cells heat inactivated / killed at 100 ° C plus the resM blend micronutrient composition. The resM micronutrient blend composition of the invention comprises chromium picolinate, white mulberry extract, fenugreek extract, vitamin B 12, and vitamin D3 as detailed in Table 2 below.TABLE 2Serving size: 1 Capsule (vegan, Hypromellose), containing: microcrystalline cellulose, magnesium stearate, silicon dioxide.EXAMPLE 3

[0069] ResM Micronutrient Blend Alone - increase in GLP-1 protein production.

[0070] As shown in Figure 1A, human gut Caco-2 cells grown with added resM micronutrient blend portion alone per Tables 1 & 2 (10 mg / mL) demonstrated a three-fold increase in GLP-1 protein production compared to control in the cell culture supernatant after co-culture 24-hour incubation at 37°C as determined by GLP-1 ELISA kit (Invitrogen, BMS2194 available from ThermoFisher Scientific, Waltham, Massachusetts) per manufacturer’s protocol.

[0071] And further, Figure 1B shows in a glucose model resM micronutrient blend portion alone per Tables 1 & 2 (10 mg / mL) demonstrated a significant increase in glucose consumption compared to control within 24 hours, demonstrating enhance metabolic activity in human gut Caco-2 cells.EXAMPLE 4

[0072] ResM™ blend of RSB11®-heat inactivated (HI) postbiotic and the resM micronutrient blend - increase in GLP-1 protein production.

[0073] As shown in Figure 2, human gut Caco-2 cells grown with the resM formulation per Table 2 which not only demonstrated a three-fold increase in GLP-1 protein production compared to control in the cell culture supernatant after co-culture 24-hour incubation at 37 ° C as determined by GLP-1 ELISA kit (Invitrogen, BMS2194) per manufacturer’s protocol, but also demonstrated a stronger, synergistic effect in the combination when compared to the components, separately. This result demonstrated robust enteroendocrine and metabolic activation using this product in human gut Caco-2 cells.EXAMPLE 5

[0074] ResM Micronutrient Blend Alone and RSB11®-heat inactivated (HI) postbiotic alone - increase in GLP-1 protein in Human Ileum Organoids.

[0075] Human organoids of the ileum are three-dimensional, self-organizing structures that mimic the small intestine's ileum region, typically using stem cells to partially recreate the tissue's identity, cell types, and functions in a lab setting. These lab-grown "mini-organs" can be used to study disease, test drug efficacy and toxicity, and gain a better understanding of gut health.

[0076] Ex vivo model

[0077] Human ileum organoids were commercially obtained (3dGRO™ Human Ileum Organoids, Age 19, Prep HT-105-I; InSphero (Brunswick, Maine) or similar source). Organoids were cultured in Matrigel within 24-well plates and maintained at 37 °C, pH 7.5-8.0. Treatments included (1) resM micronutrient blend portion alone (10 mg / mL) and (2) RSB11-HI postbioticalone (3 mg / mL). Each treatment was applied for 24 and 48 hours. After incubation, supernatants were collected for Active GLP-1 ELISA (Invitrogen BMS2194), and organoids were fixed and immunostained for GLP-1 (GFP green), ZO-1 (tight-junction marker), and DAPI (nuclei). Images were acquired by fluorescence microscopy (4X and 20X). Data represent mean ± SD of two independent experiments. Significant increases in GLP-1 protein intensity were observed, indicated by increased GFP(green) staining, confirming enteroendocrine activation in the ex vivo model.

[0078] Analysis software: GraphPad Prism 10.1.2 (One-way ANOVA, Dunnett's post-hoc test). Statistical significance: *p < 0.05: **p < 0.01: ***p < 0.001; ****p < 0.0001.

[0079] As shown in Figure 3A, human ileum organoids treated with resM micronutrient blend portion alone per Tables 1 & 2 (10 mg / mL) significantly increased active GLP-1 secretion in the ileal organoids after 24 hours compared to control. This result demonstrated robust enteroendocrine activation in a physiologically relevant ex vivo model.

[0080] As shown in Figure 3B, human ileum organoids treated with RSB11®-heat inactivated (HI) postbiotic alone per Tables 1 & 2 (3 mg / mL) significantly increased active GLP-1 secretion in the ileal organoids after 16 hours compared to control. This result demonstrated robust enteroendocrine activation in a physiologically relevant ex vivo model.

[0081] It is reasonably expected and suggested by the data that in this ex vivo model, the combination of Table 2 (and Example 4) would provide a synergistic improvement in GLP-1 secretion when compared to each component, alone when applied to human tissues such as human ileum organoids.

[0082] Furthermore, GLP-1 -positive cells were found to be localized in human ileum organoids by imaging using immunofluorescence, as disclosed herein. The GLP-1 -positive cells were found to be located within the epithelial layer, supporting functional enteroendocrine activity in human organoid models. Fluorescence protocol: Organoids were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked in 5% BSA. Primary antibodies against GLP-1 and ZO-1 were incubated overnight at 4 °C, followed by Alexa-Fluorconjugated secondary antibodies and DAPI counterstain. Images were captured under identical exposure settings to enable comparative intensity analysis.

[0083] Various combinations are contemplated as shown in the above Examples. Dosages may be varied as appropriate to age, height, and weight of the human subject.

[0084] It is expected that oral delivery of a dietary supplement, food product, or nutraceutical, comprising a blend of postbiotic compositions derived from bacterial strains selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Lactobacillus rhamnosus, as described herein would be useful in supporting metabolic health in a human subject.

[0085] For example, a blend as described above may be used to improve gastrointestinal health (thus providing relief from gas, bloating, constipation, loose stool, diarrhea, or other discomfort), and / or provide improvement in metabolic function in a human subject, for example to increase production of glucagon-like petide-1 (GLP-1) in a subject thereby improving glucose metabolism and ameliorate pre-diabetes and diabetes mellitus Type 2 (T2DM) symptoms in a human subject.EXAMPLE 6

[0086] Clinical Trial Protocol for resM micronutrient blend ± RSB 11-HI (ResB Metabolic Health Study)

[0087] A double-blind, randomized, placebo-controlled clinical trial (IRB approved, Sterling IRB #13345, Approval March 28, 2025) is ongoing to evaluate the metabolic benefits of resM micronutrient blend and RSB 11-HI formulations in overweight or mildly obese adult subjects. Subjects receive oral capsules daily for 8 weeks with follow-up at weeks 4 and 8. Primary endpoints include fasting glucose, HOMA-IR, and serum GLP-1 levels. Secondary endpoints include body composition, lipid profile, and self-reported appetite / satiety scores. Data analysis is in progress. Thus, it is expected that weight may decrease and metabolic biomarkers (such as blood glucose, hemoglobin A1C (HbAlc), and insulin) will normalize or improve compared to baseline or placebo. Other salutary changes are expected such as increases in glucagon-like peptide 1 (GLP-1) levels compared to baseline or placebo.

[0088] Test Product: resM™ GLP-1 Postbiotic, 1 Capsule Daily for 8 weeks

[0089] Study rationale

[0090] Given the functions of the ingredients, resM™ supplementation may offer an effective and tolerable approach to weight loss by enhancing gut microbiota health, regulating GLP-1 and insulin secretion, and reducing systemic inflammation. This randomized controlled trial aims to evaluate the efficacy of resM™ on primary endpoints such as weight reduction, BMI, changes in metabolic biomarkers, and reported food cravings. Exploratory endpoints includeassessing changes in GLP-1, DPP-4, depression, and gut microbiome composition via 16S sequencing. Safety will be assessed through comprehensive blood chemistry and hematological monitoring as well as adverse event reporting to ensure supplement tolerability for daily use.

[0091] This study aims to address a critical gap in metabolic health management by evaluating a postbiotic and herbal intervention designed to improve gut microbiome balance and metabolic function. Findings from this trial will help determine whether resM™ can serve as an effective supplement for individuals experiencing weight management and metabolic health challenges.

[0092] Protocol title: A Double-Blind, Randomized Placebo-Controlled Study to Evaluate a Postbiotic Supplement in Supporting Weight Loss and Metabolic Health

[0093] Study Objectives and Endpoints are shown in Table 3.TABLE 3

[0094] Safety assessments are shown in Table 4.TABLE 4

[0095] Sample size and Study population

[0096] This study will include 80 adult participants (40 active, 40 placebo) who meet the inclusion criteria for BMI and metabolic health. The ratio of resM™: Placebo groups will be 1:1.

[0097] Study Duration and Interventions

[0098] The study duration will be 56 days (8 weeks). Participants will be virtually screened for qualification up to 14 days before baseline (Day 0) to confirm eligibility. On Day 1, eligible participants will be randomized in a 1: 1 ratio to receive either resM™ or placebo and will begin the 8-week intervention. Participants will self-administer 1 capsule per day with or without food. Study reporting will include an end-of-study report at Week 8 (Day 56 ± 3 days).

[0099] Inclusion criteria[000100] 1. Provide voluntary signed and dated informed consent.[000101] 2. Be in good health as determined by medical history.[000102] 3. Age between 18 and 65 years (inclusive).[000103] 4. Body Mass Index of 25.0 - 40.0 (inclusive).[000104] 5. Subject agrees to maintain existing dietary and physical activity patterns throughout the study period.[000105] 6. Agree to refrain from other probiotic and postbiotic supplement products throughout the duration of the trial.[000106] 7. Subject is willing and able to comply with the study protocol.[000107] 8. Willing and able to agree to the requirements and restrictions of this study, be willing to give voluntary consent, be able to understand and read the questionnaires, and carry out all study related procedures.[000108] Exclusion criteria[000109] 1. History of unstable or new-onset cardiovascular / cardiorespiratory, liver, or renal conditions.[000110] 2 Alcohol abuse (more than 2 standard alcoholic drinks per day or more than 10 drinks per week) or drug abuse or dependence within the past 6 months.[000111] 3. Previous bariatric surgery.[000112] 4. Current smokers or smoking within the past month.[000113] 5. History of hyperparathyroidism or an untreated thyroid condition.[000114] 6. History of malignancy in the previous 5 years except for non-melanoma skin cancer (basal cell cancer or squamous cell cancer of the skin).[000115] 7. Other known gastrointestinal or metabolic conditions that might impact nutrient absorption or metabolism, e g., short bowel syndrome, irritable bowel syndrome (IBS), diarrheal illnesses, history of colon resection, gastroparesis, Inborn-Errors-of-Metabolism (such as PKU).[000116] 8. Chronic inflammatory condition (e.g., rheumatoid arthritis, Crohn's, ulcerative colitis, Lupus, HIV / AIDS, etc.).[000117] 9. Previous medical diagnosis of gout or fibromyalgia.[000118] 10. Pregnant women, women trying to become pregnant, women less than 120 days postpartum or nursing women. Any woman will have to take report their pregnancy status prior to enrolling and during the trial if they become pregnant.[000119] 11. Known sensitivity to any ingredient in the test formulations as listed in the product label.[000120] 12. Currently participating in another research study with an investigational product or have been in another research study in the past 30 days.[000121] Study products. Study products are defined herein as any test product or placebo intended to be administered to a study participant according to the study protocol. See, Table 5.TABLE 5[000122] Narrative of Study Procedures[000123] All screening evaluations must be completed and reviewed to confirm that potential participants meet all eligibility criteria. The investigator will maintain a screening log to record details of all participants screened and to confirm eligibility or record reasons for screening failure, as applicable.[000124] Virtual “Visit” 1: Screening (Day -14 to Day -1). On the first day of participant interaction, the overall details of the study and procedures to be undertaken will be explained to the participant. The following information will be recorded, and procedures carried out: Obtain informed consent and provide a signed copy to participant. Qualify participants:[000125] • Collect demographic information (age, sex, ethnicity, race)[000126] • Review medical history[000127] • Review / record use of concomitant medications and therapies, and dietary supplements[000128] • Calculate body mass index (BMI; kg / m2)[000129] • Review inclusion / exclusion criteria[000130] If eligible, participant will enter the study and be randomized to resM™ or placebo. ResBiotic Nutrition will mail study product and collection kits to the participant.[000131] Virtual Visit 2: Baseline (Day 0). Between Visit 1 and 2, the participant will:[000132] • Receive study product and stool collection kits in the mail[000133] • Collect first stool sample within 3 days prior to starting first dose and ship to laboratory[000134] • Collect first fasting blood draw at clinic within 3 days to starting first dose [000135] Once participants have completed the pre-baseline activities, they will perform the following:[000136] • Complete cravings questionnaire[000137] • Complete depression questionnaire[000138] • Record body weight[000139] • Calculate and record BMI[000140] • Remind the participants about study requirements in the inclusion criteria [000141] • Consume first dose of study product[000142] • Complete daily reminder survey and record adverse events as applicable [000143] Virtual Visit 3: Week 1 (Day 7±3). Between Visit 2 and 3, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 1 (Day 7±3) the following procedures will occur:[000144] • Complete cravings questionnaire[000145] • Record body weight[000146] • Calculate and record BMI[000147] Virtual Visit 4: Week 2 (Day 14±3). Between Visit 3 and 4, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 2 (Day 14±3) the following procedures will occur:[000148] • Complete cravings questionnaire[000149] • Record body weight[000150] • Calculate and record BMI[000151] Virtual Visit 5: Week 3 (Day 21±3). Between Visit 4 and 5, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 3 (Day 21±3) the following procedures will occur:[000152] • Complete cravings questionnaire[000153] • Record body weight[000154] • Calculate and record BMI[000155] Virtual Visit 6: Week 4 (Day 28±3). Between Visit 5 and 6, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 4 (Day 28±3) the following procedures will occur:[000156] • Complete cravings questionnaire[000157] • Record body weight[000158] • Calculate and record BMI[000159] Virtual Visit 7: Week 5 (Day 35±3). Between Visit 6 and 7, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 5 (Day 35±3) the following procedures will occur:[000160] • Complete cravings questionnaire[000161] • Record body weight[000162] • Calculate and record BMI[000163] Virtual Visit 8: Week 6 (Day 42±3). Between Visit 7 and 8, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 6 (Day 42±3) the following procedures will occur:[000164] • Complete cravings questionnaire[000165] • Record body weight[000166] • Calculate and record BMI[000167] Virtual Visit 9: Week 7 (Day 49±3). Between Visit 8 and 9, participants will selfadminister 1 capsule / day every day and complete daily reminder survey and record adverse events as applicable. At Week 7 (Day 49±3) the following procedures will occur:[000168] • Complete cravings questionnaire[000169] • Record body weight[000170] • Calculate and record BMI[000171] Virtual Visit 10: Week 8, End-of-Study (Day 56±3). Between Visit 9 and 10, participants will:[000172] • Self-administer 1 capsule / day every day[000173] • Complete daily reminder survey and record adverse events as applicable [000174] • Collect final stool sample within 3 days prior to taking last dose and ship to laboratory[000175] • Collect final fasting blood draw at clinic within 3 days to taking last dose [000176] Also, at Week 8 (Day 56±3) the following procedures will occur:[000177] • Complete cravings questionnaire[000178] • Complete depression questionnaire[000179] • Record body weight[000180] • Calculate and record BMI[000181] Efficacy assessments[000182] Trait Food Cravings Questionnaire (Reduced)[000183] The Trait Food Cravings Questionnaire (Reduced) (FCQ-T-r) instrument used to assess food cravings where scores have been found to be positively associated with eating pathology, body mass index (BMI), low dieting success and increases in state food craving during cognitive tasks involving appealing food stimuli. Scores will be recorded weekly via survey.[000184] Change in weight and BMI will be recorded, weekly via survey.[000185] Gut microbiome. Stool samples will be collected by participants at home at baseline and end-of-study. Samples will be shipped to a laboratory for gut microbiome analysis using 16S shallow shotgun metagenomics sequencing.[000186] Blood biomarkers[000187] Fasting blood samples will be collected at baseline and end-of-study visits to a clinic and assessed for changes in biomarkers in the blood. Blood draw tests are shown in Table 6.TABLE 6[000188] PHQ-9 Depression Questionnaire[000189] The Patient Health Questionnaire 9 (PHQ-9) is a tool for detecting depression as well as measuring its severity. Scores will be recorded at baseline and end-of-study.[000190] The use of the terms “a,” “an,” “the,” and similar referents in the context of describing the present invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. Use of the term “about” is intended to describe values either above or below the stated value in a range of approximately ±10%; in other embodiments, the values may range in value above or below the stated value in a range of approximately ±5%; in other embodiments, the “values may range in value above or below the stated value in a range of approximately ±2%; in other embodiments, the values may range in value above or below the stated value in a range of approximately ±1%. The preceding ranges are intended to be made clear by context, and no further limitation is implied. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise stated. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the invention.[000191] While in the foregoing specification this invention has been described in relation to certain embodiments thereof, and many details have been put forth for the purpose of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein can be varied considerably without departing from the basic principles of the invention.[000192] All references cited herein are incorporated by reference in their entireties. The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification, as indicating the scope of the invention.

Claims

CLAIMSWhat is claimed is:

1. A dietary supplement, food product, or nutraceutical for use in supporting metabolic health in a human subject, comprising a blend of postbiotics derived from bacterial strains selected from the group consisting of Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Lactobacillus rhamnosus.

2. The dietary supplement of claim 1, comprising postbiotics derived from probiotic bacteria, and a micronutrient and herbal extract blend comprising micronutrients chromium picolinate, and vitamins B 12 and D3, and herbal extracts comprising white mulberry extract, and fenugreek extract.

3. The dietary supplement of claim 2, wherein the postbiotics are derived from heat-killed or heat-inactivated Lactiplantibacilli or from heat-killed or heat-inactivated Lactobacilli.

4. The dietary supplement of claim 3, wherein the postbiotics are derived from Lactiplantibacillus plantarum RSB11® HI.

5. The dietary supplement of claim 4, wherein the Lactiplantibacillus plantarum RSB 11® HI is present in an amount up to 16 billion cells, the micronutrient and herbal blend is resM blend is present in an amount up to 460 mg, comprising micronutrients chromium picolinate present in an amount up to 600 mcg, vitamin B 12 is present in an amount up to 200 mcg, and vitamin D3 is present in an amount up to 10 mcg or 400 IU, and herbal extracts comprising white mulberry extract is present in an amount up to 250 mg of weight white mulberry extract, and the fenugreek extract is present in an amount up to 200 mg.

6. The dietary supplement of claim 5, wherein the composition is administered in an oral dosage form selected from tablets, capsules, solutions, suspensions, powders, gums, or confectionaries.

7. The dietary supplement of claim 5, wherein the composition is administered in a delivery form selected from sublingual dissolvable tablets dissolving under and on the tongue, liquid drops, beverages, and edible or dissolvable film dosage forms selected from hydrophilic polymers, oral dissolvable films, or oral dissolvable strips.

8. A method of modulating the gut microbiome of a human subject comprising the step of administering an effective amount of a nutraceutical composition comprising a postbiotic derived from Lactiplantibacillus plantarum, and a micronutrient blend comprising effective amounts of micronutrients comprising chromium picolinate, white mulberry extract, fenugreek extract, and vitamins B 12 and D3.

9. The method of claim 8 wherein, the modulating of the gut microbiome of a human subject results in increased production of gut hormones.

10. The method of claim 9 wherein the increase production of gut hormone is increased glucagon-like peptide 1 (GLP-1) production resulting in improved insulin homeostasis, improved glucose metabolism, and modulation to a healthy weight of the human subject.

11. The method of claim 10 wherein the effective amount of a nutraceutical composition comprises Lactiplantibacillus plantarum RSB 11® HI is present in an amount up to 16 billion cells, the micronutrient resM blend is present in an amount up to about 460 mg, the chromium picolinate is present in an amount up to about 600 mcg, the white mulberry extract is present in an amount up to about 250 mg, the fenugreek extract is present in an amount up to about 200 mg, and vitamins B 12 is present in an amount up to about 200 mcg, and vitamin D3 is present in an amount up to about 10 mcg or about 400 IU.

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