WO2022192854A1 - Méthodes d'induction de bioptérine et de métabolites apparentés - Google Patents

Méthodes d'induction de bioptérine et de métabolites apparentés Download PDF

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WO2022192854A1
WO2022192854A1 PCT/US2022/071003 US2022071003W WO2022192854A1 WO 2022192854 A1 WO2022192854 A1 WO 2022192854A1 US 2022071003 W US2022071003 W US 2022071003W WO 2022192854 A1 WO2022192854 A1 WO 2022192854A1
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lactobacillus
subject
social
reuteri
derivatives
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PCT/US2022/071003
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English (en)
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Mauro Costa-Mattioli
Shelly Alexandra BUFFINGTON
Sean DOOLING
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Baylor College Of Medicine
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system

Definitions

  • Embodiments of the disclosure concern at least the fields of cell biology, molecular biology, biochemistry, neurology, behavior, gastroenterology, and medicine.
  • the microbiome is another important source of variation (Fischbach and Segre, 2016).
  • Humans are the bearers not only of their own cells and genes, but also of the microorganisms (and their genes) living with them.
  • the phenotypes of a subject may arise from complex interactions between the genetics of the host and microorganisms, which led to the notion of the holobiont [an animal or plant host together with all associated microorganisms living in symbiosis with it (Margulis, 1993)] and the hologenome [(Rosenberg etal., 2007), the sum of all microbial genes plus the genes of the host].
  • the present disclosure satisfies a long-felt need in the art to treat neurological and neurobehavioral disorders.
  • the current disclosure fulfills the need in the art by providing methods and compositions for treating or preventing a medical condition in which at least one symptom is a social behavior deficiency, an oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels wherein the condition is a neurodevelopmental, psychiatric, and/or neurodegenerative disorder.
  • a subject in need of treatment and/or prevention of a social behavior deficiency, an oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels due to a neurodevelopmental, psychiatric, or neurodegenerative disorder of any cause is provided an effective amount of one or more bacteria species and/or one or more metabolites to treat and/or prevent one or more social behavior deficiencies, one or more oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels.
  • the treatment results in improving the social behavior deficiency, oxytocinergic system dysfunction, and/or abnormal biopertin metabolite levels
  • the treatment results in improving the social behavior deficiency, oxytocinergic system dysfunction, and/or abnormal biopertin metabolite levels to within a normal spectrum.
  • Embodiments of the disclosure include methods for treating or preventing one or more social behavior deficiencies, methods for treating or preventing one or more oxytocinergic system dysfunctions, methods for treating or preventing abnormal biopterin metabolite levels, methods of treating or preventing a neurodevelopmental disorder, methods for treating or preventing a psychiatric disorder, methods for treating or preventing a neurodegenerative disorder, methods for reducing the severity of one or more social behavior deficiencies, methods for reducing the severity of one or more oxytocinergic system dysfunctions, methods for reducing the severity of abnormal biopterin metabolite levels, methods for reducing the severity of a neurodevelopmental disorder, methods for reducing the severity of a psychiatric disorder, methods for reducing the severity of a neurodegenerative disorder, methods for determining a risk of developing one or more social behavior deficiencies, methods for determining a risk of developing one or more oxytocinergic system dysfunctions, methods for determining a risk of developing of abnormal biopterin metabolite levels, methods for determining
  • Methods of the present disclosure can include at least 1, 2, 3, 4, 5, or more of the following steps: administering one or more bacterial compositions to a subject, administering one or more metabolite compositions to a subject, administering one or more compositions comprising one or more bacteria and one or more metabolites to a subject, determining a subject to have a higher risk of developing one or more social behavior deficiencies, determining that one or more social behavior deficiencies pose a greater risk to the health or life of the subject, determining a subject to have a higher risk of developing one or more oxytocinergic system dysfunctions, determining that one or more oxytocinergic system dysfunctions pose a greater risk to the health or life of the subject, determining a subject to have a higher risk of developing abnormal biopterin metabolite levels, determining that abnormal biopterin metabolite levels pose a greater risk to the health or life of the subject, determining a subject to have a higher risk of developing a neurodevelopmental disorder, determining that the neurodevelopmental disorder poses
  • compositions of the present disclosure can include at least 1, 2, 3, or more of the following components: a bacterial composition or formulation, a metabolite composition or formulation, a probiotic, a prebiotic, and a pharmaceutical excipient. It is contemplated that any one or more of these components may be excluded from certain embodiments of the disclosure.
  • the bacteria is from the Lactobacillus genus.
  • the Lactobacillus species is L. reuteri.
  • the L. reuteri may be delivered as a single species or as one of multiple species of non-pathogenic bacteria.
  • the Lactobacillus species is one or more L. reuteri strains.
  • the Lactobacillus species is L. reuteri strain 6475.
  • the Lactobacillus species is L. reuteri strain 6475 and other L. reuteri strains.
  • the metabolite is a metabolite of tetrahydrobiopterin (BH4) or one or more derivatives thereof.
  • BH4 tetrahydrobiopterin
  • BH4 or one or more derivatives thereof in the subject treats or prevents the one or more social behavior deficiencies
  • treating or preventing autism spectrum disorder in a subject by increasing levels of tetrahydrobiopterin (BH4) or one or more derivatives thereof in the subject wherein increasing levels of BH4 or one or more derivatives thereof in the subject treats or prevents the autism spectrum disorder
  • BH4 or one or more derivatives thereof in the subject treats or prevents the oxytocinergic system dysfunction
  • BH4 or one or more derivatives thereof in the subject treats or prevents the cardiovascular disease
  • methods of treating or preventing cardiovascular disease in a subject by increasing levels of BH4 or one or more derivatives thereof in the subject wherein increasing levels of BH4 or one or more derivatives thereof in the subject treats or prevents the cardiovascular disease
  • treating or preventing cardiovascular disease in a subject having or at risk of having cardiovascular disease by increasing levels of BH4 or one or more derivatives thereof in the gut of the subject wherein increasing levels of BH4 or one or more derivatives thereof in the gut of the subject treats or prevents the cardiovascular disease.
  • BH4-deficient hyperphenylalaninemia and/or tyrosinemia in some aspects, are methods of treating or preventing BH4-deficient hyperphenylalaninemia and/or tyrosinemia in a subject by increasing levels of BH4 or one or more derivatives thereof in the subject, wherein increasing levels of BH4 or one or more derivatives thereof in the subject treats or prevents the BH4-deficient hyperphenylalaninemia and/or tyrosinemia; and treating or preventing BH4-deficient hyperphenylalaninemia and/or tyrosinemia in a subject having or at risk of having BF14 -deficient hyperphenylalaninemia and/or tyrosinemia by increasing levels of BH4 or one or more derivatives thereof in the gut of the subject, wherein increasing levels of BH4 or one or more derivatives thereof in the gut of the subject treats or prevents the BH4-deficient hyperphenylalaninemia and/or tyrosinemia.
  • BH4 metabolism pathway in a subject by administering to the subject a therapeutically effective amount of Lactobacillus reuteri and/or a bioactive extract or fraction thereof, thereby stimulating the BH4 metabolism pathway in the subject; increasing levels of BH4 or one or more derivatives thereof in a subject by administering to the subject a therapeutically effective amount of Lactobacillus reuteri and/or a bioactive extract or fraction thereof, thereby increasing the levels of BH4 or one or more derivatives thereof in the subject; and increasing levels of BH4 or one or more derivatives thereof in the gut of a subject having or at risk of having neurodevelopmental, neurodegenerative, psychiatric disorders, cardiovascular, and/or metabolic disorders by administering a therapeutically effective amount of Lactobacillus reuteri and/or a bioactive extract or fraction thereof, wherein administering the therapeutically effective amount of the Lactobacillus reuteri and/or a bioactive extract
  • a formulation comprising Lactobacillus reuteri and/or a bioactive extract or fraction thereof for increasing levels of one or more BH4 or one or more derivatives thereof in a subject; use of Lactobacillus reuteri and/or a bioactive extract or fraction thereof for stimulating a BH4 metabolism pathway in a subject; use of Lactobacillus reuteri and/or a bioactive extract or fraction thereof in the manufacture of a medicament for increasing levels of BH4 or one or more derivatives thereof in a subject; and use of Lactobacillus reuteri and/or a bioactive extract or fraction thereof in the manufacture of a medicament for stimulating a BH4 metabolism pathway in the subject.
  • the one or more BH4 derivatives comprise BH4-4a-carbinolamine tetrahydrobiopterin, dihydrobiopterin (BH2), quinonoid-BH2, or biopterin. In some embodiments, the one or more BH4 derivatives comprise biopterin.
  • the levels of BH4 or one or more derivatives thereof are increased by administering a therapeutically effective amount of a formulation comprising Lactobacillus reuteri and/or a bioactive extract or fraction thereof.
  • the Lactobacillus reuteri comprises Lactobacillus reuteri strains ATCC PTA 6475.
  • ATCC PTA 6475 (MM4-1A), ATCC 6A15::pocR, ATCC PTA 4659 (MM2-3), ATCC PTA 5289 (FJ1), CRL1324, 1048, 1063, 1073, 173.5, 27.4, atcc55739, atcc53608, jw2015, jw2019, ks6, lem83, lr85573, tmwll294, 10c2, p97, pg3b, 32, 676, cp447, 6sl5, cp415, lpl67.67, lpal, tmwll46, tmwll37, 100.93, ad23, dbc2, 100-23, dsm20053, ilc4, mlc3, mouse56, n2j, r21c, 11283, 11284, kel, ky21, mfl4c, mf2-3, mf23, tl, 1204, 1366, at
  • the Lactobacillus reuteri comprises Lactobacillus reuteri strains ATCC PTA 6475, DSM17938, ATCC PTA4659, ATCC PTA 5289, ATCC 55730, CRL1324, DSM20016, CF4-6G, ATCC55730, CF48-3A1, M27U15, or a combination thereof.
  • the Lactobacillus reuteri comprises Lactobacillus reuteri strain ATCC PTA 6475.
  • the formulation further comprises an effective amount of one or more of prebiotics, nutrients, yeast, additional bacteria species, or a combination thereof.
  • the one or more prebiotics comprise simple sugars, complex carbohydrates, fibers, fats, proteins, vitamins, minerals, polyphynols, inulin, arabinoxylan, xylose, soluble fiber dextran, soluble com fiber, polydextrose, lactose, N- acetyl-lactosamine, glucose, galactose, fructose, rhamnose, mannose, uronic acids, 3’- fucosyllactose, 3’-sialylactose, 6’-sialyllactose, lacto-N-neotetraose, 2’-2’-fucosyllactose, trans- galactooligosaccharides, glucooligosaccharides, isomaltooligo
  • the one or more nutrients comprise one or more of vitamins, lipids, proteins, amino acids, nucleotides, peptides, cofactors, xenobiotics, carbohydrates, or a combination thereof.
  • the one or more yeast or additional bacteria species comprise Lactobacillus acidophilus, Lactobacillus bifidus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus vaginalis, Akkermansia muciniphilus , Bacillus subtlis
  • lactis Bifidobacterium longum, Bifidobacterium pseudolongum, Collinsella unclassified, Enterococcus faecalis, Enterococcus faecium, Helicobacter hepaticus, Lactococcus lactis, Olsenella unclassified, Parabacteroides distasonis, Streptococcus thermophilis , Saccharomyces boulardii, or a combination thereof.
  • the subject is given an effective amount of one or more additional therapies to treat or prevent one or more social behavioral deficiencies in the subject.
  • the one or more additional therapies comprise one or more of prebiotics, nutrients, yeast, additional bacteria species other than L. reuteri, and/or behavioral therapy.
  • the one or more prebiotics comprise simple sugars, complex carbohydrates, fibers, fats, proteins, vitamins, minerals, polyphynols, inulin, arabinoxylan, xylose, soluble fiber dextran, soluble com fiber, polydextrose, lactose, N- acetyl-lactosamine, glucose, galactose, fructose, rhamnose, mannose, uronic acids, 3’- fucosyllactose, 3’-sialylactose, 6’-sialyllactose, lacto-N-neotetraose, 2’-2’-fucosyllactose, trans- galactooligosaccharides, glucooligosaccharides, isomaltooligosaccharides, lactosucrose, polydextrose, soybean oligosaccharides, arabinose, cellobiose, fructose, fucose, gal
  • the one or more nutrients comprise one or more of vitamins, lipids, proteins, amino acids, nucleotides, peptides, cofactors, xenobiotics, carbohydrates, or a combination thereof.
  • the one or more yeast or additional bacteria species comprise Lactobacillus acidophilus, Lactobacillus bifidus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus iners, Lactobacillus jensenii, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus vaginalis, Akkermansia muciniphilus, Bacillus subtlis,
  • lactis Bifidobacterium longum, Bifidobacterium pseudolongum, Collinsella unclassified, Enterococcus faecalis, Enterococcus faecium, Helicobacter hepaticus, Lactococcus lactis, Olsenella unclassified, Parabacteroides distasonis, Streptococcus thermophilis, Saccharomyces boulardii, or a combination thereof.
  • the formulation and/or one or more additional therapies is delivered via a pill or suppository.
  • the formulation and/or one or more additional therapies is a food, beverage, yogurt, dry powder, or dry powder mixed in water or another beverage.
  • the formulation and/or one or more additional therapies is administered to the subject by an oral route, a rectal route, a nasal route, by naso/oro-gastric gavage, or by a combination thereof.
  • the subject when providing the formulation to the subject, had an altered gut microbiome.
  • the subject is identified as having an altered gut microbiome prior to provision of the Lactobacillus reuteri , formulations comprising Lactobacillus reuteri , and/or one or more additional therapies to the subject to treat or prevent any of the deficiencies, dysfunctions, and/or disorders disclosed herein.
  • the social behavioral deficiency comprises impaired sociability, preference for social novelty, difficulty in social use of verbal and nonverbal communication, or a combination thereof.
  • the subject has a neurodevelopment disorder.
  • the subject has autism spectrum disorder or other neurological disorders associated with social dysfunction.
  • the oxytocinergic system dysfunction comprises social behavior deficits, social cognitive and/or memory deficits, addictive behavior, anxiety and/or depressive behavior, abnormal feeding behavior, or a combination thereof.
  • the subject has or is at risk of having one or more of William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease.
  • the subject has or is at risk of having cardiovascular disease. In some embodiments, the subject has or is at risk of having BH4-deficient hyperphenylalaninemia and/or tyrosinemia.
  • “Individual, “subject,” and “patient” are used interchangeably and can refer to a human or non-human. [0029] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the measurement or quantitation method.
  • A, B, and/or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.
  • A, B, and/or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C.
  • “and/or” operates as an inclusive or.
  • compositions and methods for their use can “comprise,” “consist essentially of,” or “consist of’ any of the ingredients or steps disclosed throughout the specification. Compositions and methods “consisting essentially of’ any of the ingredients or steps disclosed limits the scope of the claim to the specified materials or steps which do not materially affect the basic and novel characteristic of the claimed disclosure.
  • the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that embodiments described herein in the context of the term “comprising” may also be implemented in the context of the term “consisting of’ or “consisting essentially of.”
  • compositions may be employed based on any of the methods described herein.
  • Other embodiments are discussed throughout this application. Any embodiment discussed with respect to one aspect of the disclosure applies to other aspects of the disclosure as well and vice versa.
  • any step in a method described herein can apply to any other method.
  • any method described herein may have an exclusion of any step or combination of steps.
  • the embodiments in the Example section are understood to be embodiments that are applicable to all aspects of the technology described herein.
  • FIG. 1A-1I Cntnap2-/- mice generated from isolated breeding lines exhibit impaired social behavior, hyperactivity, and a distinct microbiome composition compared to WT controls.
  • FIG. 1A Breeding scheme for the isolated lines ( Cntnap2 +/+ : WT-I; Cntnap2 / KO-I).
  • FIG. IB Experimental design for animals from isolated lines.
  • FIGs. 2A-2B Assays used to assess social behavior and locomotor activity in genetic mouse model for neurodevelopmental disorders.
  • FIG. 2A Scheme of 3-chamber test for social behavior and activity levels.
  • FIG. 2B Scheme of reciprocal social interaction test.
  • FIGs. 3A-3B Normal olfaction in a mouse model for a neurodevelopmental disorder.
  • FIG. 3A Scheme of the buried food test used to measure olfaction.
  • FIGs. 4A-4G KO-I mice exhibit alterations in gut microbiome composition compared to WT-I mice.
  • FIGs. 4C-4D FIGs. 4C-4D.
  • FIG. 4E Volcano plot of ASV differential abundances colored at the class taxonomic level, with points above the dashed line (representing the FDR-adjusted P-value threshold) considered statistically significant.
  • FIG. 4F Volcano plot of ASV differential abundances colored at the class taxonomic level, with points above the dashed line (representing the FDR-adjusted P-value threshold) considered statistically significant.
  • FIG. 4G Pie chart reflecting significant differences (FDR-adjusted P-value ⁇ 0.05) between groups in predicted MetaCyc gene pathway abundances.
  • FIGs. 5A-5F Oxytocin rescues social deficits, but not hyperactivity, in the Cntnap2 mouse model for neurodevelopmental disorders.
  • FIG. 5A Scheme of experimental design for oxytocin treatment studies.
  • FIG. 5A-5F Oxytocin rescues social deficits, but not hyperactivity, in the Cntnap2 mouse model for neurodevelopmental disorders.
  • FIG. 5A Scheme of experimental design for oxytocin treatment studies.
  • FIGs. 5B-5C Social behavior in KO-I mice treated with
  • FIGs. 7A-7D L. reuteri treatment increases hypothalamic oxytocin levels in the Cntnap2 / mouse model for neurodevelopmental disorders.
  • FIG. 7A Scheme of experimental design for L. reuteri treatment and immuno staining.
  • FIG. 7B Representative images of immunoreactivity in the paraventricular nucleus of the hypothalamus (Oxytocin, Green; NeuN, Red; DAPI, Blue).
  • FIG. 8A-8H L. reuteri rescues deficits in social behavior and related changes in synaptic transmission without altering locomotor activity levels in Cntnap2 / mice.
  • FIG. 8A Scheme of experimental design for the treatment with L. reuteri.
  • FIG. 8F Scheme of experimental design for electrophysiological studies.
  • FIGs. 9A-9C Treatment with L. reuteri during adulthood rescues the social deficits in the Cntnap2 / mouse model for neurodevelopmental disorders.
  • FIG. 9A Scheme of experimental design for L. reuteri treatment during adulthood.
  • FIG. 10A-10D Treatment with L. reuteri fails to rescue deficits in synaptic transmission in the medial prefrontal cortex (mPFC) of Cntnap2 mice.
  • FIG. 10A-10D Treatment with L. reuteri fails to rescue deficits in synaptic transmission in the medial prefrontal cortex (mPFC) of Cntnap2 mice.
  • mPFC medial prefrontal cortex
  • FIG. 10A Scheme of brain area targeted for electrophysiological recordings.
  • FIG. IOC Miniature excitatory post- synaptic currents
  • FIG. 10D Representative traces of mEPSCs.
  • FIGs. 11A-11L L. reuteri selectively increases metabolites from the tetrahydrobiopterin (BH4) synthesis pathway and direct BH4 treatment reverses the social deficits (but not locomotor activity levels) in Cntnap2 mice.
  • FIG. 11B Top 30 most discriminatory fecal metabolites between social (WT-I + Vehicle and KO-I + L.
  • FIG. HE Scheme of experimental design for BH4 treatment studies.
  • FIG. 11H Scheme of experimental design for electrophysiological studies.
  • FIGs. 12A-12E Tetrahydrobiopterin treatment rescues social behavior deficits but has no effect on the activity levels of GF mice.
  • FIG. 12A Scheme of experimental design for GF-BH4 studies.
  • FIG. 12A Scheme of experimental design for GF-BH4 studies.
  • FIGs. 13A-13F Blocking tetrahydrobiopterin synthesis via inhibition of sepiapterin reductase with SPRi3 prevents L. reuteri from rescuing social behavior deficits in Cntnap2 / mice.
  • FIG. 13A Scheme of experimental design for L. reuteri + SPRi3.
  • FIGs. 14A-14F Blocking tetrahydrobiopterin synthesis via inhibition of sepiapterin reductase with SPRi3 leads to social behavior deficits in WT mice.
  • FIG. 14A Scheme of experimental design for SPRi3 treatment.
  • FIG. 14A Scheme of experimental design for SPRi3 treatment.
  • FIGs. 15A-15B Oxytocin receptor antagonist prevents the rescue of social deficits by tetrahydrobiopterin in the Cntnap2 mouse model for neurodevelopmental disorders.
  • FIG. 15A Scheme of experimental design for oxytocin receptor antagonist (L-371,257) experiments.
  • the present disclosure relates to methods and compositions for treating or preventing one or more social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels in a subject or medical conditions wherein one or more social behavior deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels are at least one symptom.
  • the subject has a neurodevelopmental disorder, a psychiatric disorder, a neurodegenerative disorder, a cardiovascular disorder, and/or a metabolic disorder.
  • the subject has autism spectrum disorder (ASD), for example.
  • ASSD autism spectrum disorder
  • the subject has William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, bipolar disorder, schizophrenia, sleep disorder, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease.
  • the subject has cardiovascular disease.
  • the subject has BH4-deficient hyperphenylalaninemia and/or tyrosinemia.
  • a subject in need thereof is provided an effective amount of one or more particular bacteria and/or metabolites for a therapeutic or preventative purpose.
  • composition and activity of the gut microbiome is analyzed or measured or determined or evaluated for a subject, and that analysis may occur by any suitable method.
  • a therapeutically effective amount of a composition comprising one or more bacteria and/or bioactive extracts or fractions thereof and/or one or more metabolites is prophylactically utilized to treat, prevent, or reduce the severity of social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels in a subject having an altered gut microbiome and having a neurodevelopmental disorder, a psychiatric disorder, a neurodegenerative disorder, a cardiovascular disorder, and/or a metabolic disorder.
  • prior analysis of the composition and activity of the gut microbiome allows determination of which individuals would need the composition comprising one or more bacteria and/or bioactive extracts or fractions thereof and/or one or more metabolites, allowing for informed clinical decision making.
  • Embodiments of the disclosure concern compositions comprising an effective amount of the metabolite tetrahydrobiopterin (BH4) or one or more derivatives thereof for treatment or prevention of a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels, including in some cases a neurodevelopmental disorder, a psychiatric disorder, a neurodegenerative disorder, a cardiovascular disorder, a metabolic disorder, or other discorders characterized by abnormal biopterin metabolite levels.
  • BH4 metabolite tetrahydrobiopterin
  • a subject that has a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels or that is at risk for having a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels is provided an effective amount of a composition comprising BH4 or one or more derivatives thereof.
  • the one or more derivatives of BH4 comprise BH4-4a- carbinolamine, dihydrobiopterin (BH2), quinonoid-BH2, and/or biopterin.
  • the BH4 derivative is BH4-4a-carbinolamine.
  • the BH4 derivative is BH2.
  • the BH4 derivative is quinonoid-BH2.
  • the BH4 derivative is biopterin.
  • Biopterin is defined as the pteridine analogue in which the heterocyclic ring is substituted with amino, carbonyl oxygen and 1,2-dihydroxypropyl at the 2, 4 and 6 positions, respectively.
  • Pterins are pterin molecules, which comprise a heterocyclic compound composed of a pteridine ring system with a keto group and an amino group on positions 4 and 2 respectively, with additional substituents.
  • the substituted biopterin ring exists as fully oxidized biopterin, partially reduced dihydrobiopterin (BH2), or fully reduced BH4 forms, with BH4-4a- carbinolamine and quinonoid-BH2 intermediates formed during catalysis.
  • BH4 is oxidized to form the intermediate BH4-4a-carbinolamine.
  • BH4-4a- carbinolamine is oxidized to form the quinonoid dihydrobiopterin intermediate quinonoid-BH2.
  • quinonoid-BH2 is oxidized to form biopterin.
  • biopterins including BH4 function as endogenous enzyme cofactors for aromatic amino acid hydroxylases (AAAH), which are involved in the synthesis of a number of neurotransmitters including dopamine, norepinephrine, epinepherine, and serotonin, along with several trace amines.
  • AAAH aromatic amino acid hydroxylases
  • BH4 is the endogenous cofactor for AAAH enzymes.
  • BH4 is a cofactor for the following enzymes: TPH (tryptophan hydroxylase), which converts the amino acid tryptophan into 5-HTP (5-hydroxytryptophan), a precursor to serotonin; PAH (phenylalanine dehydroxylase), which converts the amino acid L-phenylalanine into L-tyrosine, as part of the formation of dopamine; and TH (Tyrosine hydroxylase), which converts L-tyrosine (formed in the preceding PAH reaction) into L-DOPA, a precursor to dopamine.
  • TPH tryptophan hydroxylase
  • PAH phenylalanine dehydroxylase
  • TH Tyrosine hydroxylase
  • Dopamine is a precursor for the neurotransmitters norepinephrine and epinephrine.
  • Serotonin is the precursor for the hormone melatonin.
  • BH4 a deficiency of BH4 can lead to systemic deficiencies of serotonin, dopamine, norepinephrine, and epinephrine.
  • compositions of BH4 or one or more derivatives thereof may or may not be tailored to address any deficiency in a subject’s gut microbiome or to enhance a subject’s gut microbiome.
  • the compositions may be given to a subject without having prior analysis of their gut microbiome.
  • the compositions of BH4 or one or more derivatives thereof may comprise any one or more of BH4 and/or derivatives thereof associated with efficacious therapy to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels.
  • the subject may be given one or more compositions of BH4 and/or derivatives thereof, including compositions that comprise one or more of BH4 and/or derivatives thereof that overcome any deficiencies in the subject’s gut microbiome.
  • the BH4 and/or derivatives thereof may be given to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels and/or enhance therapy to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels.
  • compositions of BH4 and/or derivatives thereof can be administered alone or in combination with one or more additional one or more additional probiotic, prebio tic, neutraceutical, or therapeutic agents disclosed herein.
  • Administration “in combination with” one or more additional therapeutic agents includes both simultaneous (at the same time) and consecutive administration in any order.
  • the compositions of BH4 and/or derivatives thereof and one or more additional therapeutic agents can be administered in one composition, or simultaneously as two separate compositions, or sequentially. Administration can be chronic or intermittent, as deemed appropriate by the supervising practitioner, including in view of any change in any undesirable side effects.
  • An effective amount of BH4 and/or derivatives thereof may be provided to a subject in need thereof and may or may not be provided with one or more additional probiotic, prebio tic, neutraceutical, or therapeutic agents.
  • the additional probiotic, prebiotic, neutraceutical, or therapeutic agents may be in the same composition as the BH4 and/or derivatives thereof, or the additional probiotic, prebiotic, neutraceutical, or therapeutic agents may be in a different composition as the BH4 and/or derivatives thereof.
  • the additional probiotic, prebiotic, neutraceutical, or therapeutic agents may or may not be given to the subject at the same time as the BH4 and/or derivatives thereof.
  • the additional probiotic, prebiotic, neutraceutical, or therapeutic agents may assist in treating and/or preventing at least one social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite level and/or the additional probiotic, prebiotic, neutraceutical, or therapeutic agents may be useful for treating and/or preventing at least one symptom of another medical condition.
  • Other medical conditions that may be treated and/or prevented at the same time with other probiotic, prebiotic, neutraceutical, or therapeutic agents include at least anxiety, cognition, schizophrenia, bipolar disorder, mood disorder, seasonal affective disorder, irritable bowel syndrome, inflammatory bowel disease (IBD), infectious diarrhea (caused by viruses, bacteria, or parasites), antibiotic-related diarrhea, skin conditions (such as eczema), urinary and vaginal conditions, allergies and colds, and oral conditions.
  • IBD inflammatory bowel disease
  • infectious diarrhea caused by viruses, bacteria, or parasites
  • antibiotic-related diarrhea skin conditions (such as eczema)
  • urinary and vaginal conditions such as eczema
  • allergies and colds and oral conditions.
  • the BH4 and/or derivatives thereof is administered prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents. In some embodiments, the BH4 and/or derivatives thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • At least 1, 2, 3, 4, 5, 6, or 7 doses (or any derivable range therein) of the BH4 and/or derivatives thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the BH4 and/or derivatives thereof is administered after the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the BH4 and/or derivatives thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) after the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • at least 1, 2, 3, 4, 5, 6, or 7 doses (or any derivable range therein) of the BH4 and/or derivatives thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) after the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • compositions comprising the BH4 and/or derivatives thereof are provided.
  • the compositions may include a pharmaceutically acceptable carrier, diluent and/or excipient, in some cases.
  • the compositions comprising BH4 and/or derivatives thereof are formulated to target a region in the gastrointestinal tract, including any portion of the gastrointestinal tract.
  • the compositions comprising BH4 and/or derivatives thereof are formulated to be delivered to any portion of the gastrointestinal tract.
  • compositions comprising BH4 and/or derivatives thereof encompassed herein may be administered in a variety of ways known or available to those skilled in the art.
  • the composition is directly or indirectly delivered to the digestive tract of the subject.
  • the compositions comprising BH4 and/or derivatives thereof are administered to the subject by a route selected from the group consisting of oral, rectal ( e.g ., by enema), nasal, and via naso/oro-gastric gavage.
  • compositions comprising BH4 and/or derivatives thereof are delivered to the subject in a form of a liquid, foam, cream, spray, powder, or gel.
  • the compositions comprising BH4 and/or derivatives thereof comprise a buffering agent, along with preservatives, stabilizers, binders, compaction agents, lubricants, dispersion enhancers, disintegration agents, antioxidants, flavoring agents, sweeteners, and coloring agents.
  • the composition can administered alone or in combination with a carrier, such as a pharmaceutically acceptable carrier or a biocompatible scaffold.
  • a carrier such as a pharmaceutically acceptable carrier or a biocompatible scaffold.
  • formulations of the composition comprise an ingestible carrier, which may be a pharmaceutically acceptable carrier such as a capsule, tablet or powder.
  • the ingestible carrier is a food product such as acidified milk, yogurt (frozen or non-frozen), milk powder, milk concentrate, cheese spreads, dressings and/or beverages.
  • the composition further comprises a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element, for example.
  • the formulation of the composition may further comprise an adjuvant, a drug, a biological compound, or a mixture thereof.
  • the formulation of the composition may be a food stuff or a medicament, in certain embodiments.
  • oral formulations include such normally employed excipients such as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. These formulations take the form of solutions, suppositories, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain about 10% to about 95% of active ingredient, preferably about 25% to about 70%.
  • the compositions are formulated for oral administration.
  • Oral administration may be achieved using a chewable formulation, a dissolving formulation, an encapsulated/coated formulation, a multi-layered lozenge (to separate active ingredients and/or active ingredients and excipients), a slow release/timed release formulation, or other suitable formulations known to persons skilled in the art.
  • the word “tablet” is used herein, the formulation may take a variety of physical forms that may commonly be referred to by other terms, such as lozenge, pill, capsule, or the like.
  • compositions of the present disclosure may be formulated for oral administration, other routes of administration can be employed, however, including, but not limited to, subcutaneous, intramuscular, intradermal, transdermal, intraocular, intraperitoneal, mucosal, vaginal, rectal, and intravenous.
  • the disclosed compositions are formulated as a capsule.
  • the capsule may be a hollow, generally cylindrical capsule formed from various substances, such as gelatin, cellulose, carbohydrate or the like.
  • compositions are formulated as a suppository.
  • the suppository may include but is not limited to the bacteria and one or more carriers, such as polyethylene glycol, acacia, acetylated monoglycerides, camuba wax, cellulose acetate phthalate, com starch, dibutyl phthalate, docusate sodium, gelatin, glycerin, iron oxides, kaolin, lactose, magnesium stearate, methyl paraben, pharmaceutical glaze, povidone, propyl paraben, sodium benzoate, sorbitan monoleate, sucrose talc, titanium dioxide, white wax and coloring agents.
  • carriers such as polyethylene glycol, acacia, acetylated monoglycerides, camuba wax, cellulose acetate phthalate, com starch, dibutyl phthalate, docusate sodium, gelatin, glycerin, iron oxides, kaolin, lactose, magnesium stearate,
  • the disclosed compositions are prepared as a tablet.
  • the tablet may include the bacteria and one or more tableting agents, such as dibasic calcium phosphate, stearic acid, croscarmellose, silica, cellulose and cellulose coating.
  • the tablets may be formed using a direct compression process, though those skilled in the art will appreciate that various techniques may be used to form the tablets.
  • the compositions comprising BH4 and/or derivatives thereof and related methods of the present disclosure utilize particular formulation methods.
  • the carrier may further comprise a disintegrant, a glidant, and/or a lubricant, such as is described in U.S. Patent 9,084,434 (incorporated by reference herein in its entirety), for example, to facilitate having a greater shelf life and/or half-life of the formulation.
  • the disintegrant may be any suitable disintegrant such as, for example, a disintegrant selected from the group consisting of sodium croscarmellose, crospovidone, gellan gum, hydroxypropyl cellulose, starch, and sodium starch glycolate.
  • the glidant may be any suitable glidant such as for example, a glidant selected from the group consisting of silicon dioxide, colloidal silicon dioxide, and talc.
  • the lubricant may be any suitable lubricant such as for example, a lubricant selected from the group consisting of calcium stearate, magnesium stearate, stearic acid, sodium stearyl fumerate, and vegetable based fatty acids.
  • the carrier is present in the composition in a range of approximately 30% w/w to approximately 98% w/w; this weight percentage is a cumulative weight percentage taking into consideration all ingredients present in the carrier.
  • the composition of the present invention may be an oral dosage form, a powder that is mixed into a liquid, or a chewing gum.
  • the oral dosage form may be selected from the group consisting of tablets, caplets, and capsules, wherein the tablets and caplets may be solid or chewable.
  • the composition is a powder, it may be mixed into a liquid that is selected from the group consisting of water, milk, juice, and yogurt.
  • the gum may be soft gum or hard chewing gum tablets.
  • probiotic species such as mannitol, sorbitol, alone or together with the additional sugar alcohol lactitol and/or a phytonutrient, such as oligomeric proanthocyanidins (OPC)
  • sugar alcohols such as mannitol, sorbitol
  • a phytonutrient such as oligomeric proanthocyanidins (OPC)
  • the formulation of the compositions comprising BH4 and/or derivatives thereof comprises one or more excipients.
  • excipients that may be used to formulate appropriate dosage forms include binders, disintegrants, lubricants, coatings, plasticizers, compression agents, wet granulation agents, and sweeteners, all of which are known to those of ordinary skill in the art to which the invention pertains. All of the following examples are provided by way of illustration and not limitation.
  • Binders are used where appropriate to help the dosage form ingredients still together. Examples of binders include carbopol, povidone, and xanthan gum.
  • Lubricants are generally always used in the manufacture of dosage forms by direct compression in order to prevent the compacted powder mass from sticking to the equipment during the tabletting or encapsulation process.
  • examples of lubricants include calcium stearate, magnesium stearate, stearic acid, sodium stearyl fumerate, and vegetable based fatty acids.
  • Disintegrants aid in the break-up of the compacted mass when placed in a fluid environment.
  • disintegrants include sodium croscarmellose, crospovidone, gellan gum, hydroxypropyl cellulose, starch, and sodium starch glycolate. Coatings are used to control the solubility of the drug.
  • coatings include carrageenan, cellulose acetate phthalate, ethylcellulose, gellan gum, matodextrin, methacrylates, methylcellulose, microcrystalline cellulose, and shellac.
  • Plasticizers are used to control the release rate of the drug from the dosage form. Examples of plasticizers include citrate esters, dibutyl sebacate, diethyl phthalate, polyvinylacetate phthalate, and triacetin.
  • Compression agents include calcium carbonate, dextrose, fructose, guar gum, honey, lactose, maltodextrin, maltose, mannitol, microcrystalline cellulose, molasses, sorbitol, starch, and sucrose.
  • Wet granulation agents include calcium carbonate, lactose, maltodextrin, mannitol, microcrystalline cellulose, povidone, and starch.
  • Sweeteners include aspartame, dextrose, fructose, honey, lactose, maltodextrin, maltose, mannitol, molasses, monoammonium glycyrrhizinate, sorbitol, sucralose, and sucrose.
  • Excipients that are generally used in the manufacture of chewable tablets include by way of illustration and not limitation, dextrose, fructose, guar gum, lactose, maltodextrin, maltose, mannitol, microcrystalline cellulose, and sorbitol. As is evident from the foregoing list, many of the same ingredients may be used for various different purposes in various different dosage forms.
  • the agents of the disclosure may be administered by the same route of administration or by different routes of administration.
  • the prebiotic is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally.
  • the microbial composition is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally.
  • the appropriate dosage may be determined based on the type of disease to be treated, severity and course of the disease, the clinical condition of the subject, the subject’s clinical history and response to the treatment, and the discretion of the attending physician.
  • the treatments may include various “unit doses.”
  • Unit dose is defined as containing a predetermined-quantity of the therapeutic composition calculated to produce the desired responses discussed above in association with its administration, i.e., the appropriate route and regimen.
  • the quantity to be administered, and the particular route and formulation, is within the skill of determination of those in the clinical arts and depends on the result and/or protection desired.
  • a unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time.
  • a unit dose comprises a single administrable dose.
  • the quantity of BH4 or one or more metabolites thereof to be administered, both according to number of treatments and unit dose depends on the treatment effect desired.
  • an effective dose is understood to refer to an amount necessary to achieve a particular effect.
  • doses in the range from 10 mg/kg to 200 mg/kg can affect the protective capability of these agents.
  • doses include doses of about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, and 200, 300, 400, 500, 1000 pg/kg, mg/kg, pg/day, or mg/day or any range derivable therein.
  • the effective dose of the pharmaceutical composition is one which can provide a blood level of about 1 pM to 150 pM.
  • the effective dose provides a blood level of about 4 pM to 100 pM.; or about 1 pM to 100 pM; or about 1 pM to 50 pM; or about 1 pM to 40 pM; or about 1 pM to 30 pM; or about 1 pM to 20 pM; or about 1 pM to 10 pM; or about 10 pM to 150 pM; or about 10 pM to 100 pM; or about 10 pM to 50 pM; or about 25 pM to 150 pM; or about 25 pM to 100 pM; or about 25 pM to 50 pM; or about 50 pM to 150 pM; or about 50 pM to 100 pM (or any range derivable therein).
  • the dose can provide the following blood level of the agent that results from a therapeutic agent being administered to a subject: about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
  • the therapeutic agent that is administered to a subject is metabolized in the body to a metabolized therapeutic agent, in which case the blood levels may refer to the amount of that agent.
  • the blood levels discussed herein may refer to the unmetabolized therapeutic agent.
  • dosage units of pg/kg or mg/kg of body weight can be converted and expressed in comparable concentration units of pg/ml or mM (blood levels), such as 4 mM to 100 pM. It is also understood that uptake is species and organ/tissue dependent. The applicable conversion factors and physiological assumptions to be made concerning uptake and concentration measurement are well-known and would permit those of skill in the art to convert one concentration measurement to another and make reasonable comparisons and conclusions regarding the doses, efficacies and results described herein.
  • compositions are administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically or prophylactically effective for the subject being treated.
  • Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are peculiar to each individual. Suitable regimes for initial administration and boosters are also variable, but are typified by an initial administration followed by subsequent administrations. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability and toxicity of the particular therapeutic substance or other therapies a subject may be undergoing.
  • compositions will be pharmaceutically acceptable or pharmacologically acceptable.
  • pharmaceutically acceptable or “pharmacologically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal, or human.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in immunogenic and therapeutic compositions is contemplated.
  • Embodiments of the disclosure concern compositions comprising an effective amount of one or more probiotics for treatment or prevention of a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels, including in some cases a neurodevelopmental disorder, a psychiatric disorder, a neurodegenerative disorder, a cardiovascular disorder, and/or a metabolic disorder.
  • a subject that has a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels or that is at risk for having a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels is provided an effective amount of a composition comprising one or more probiotics.
  • probiotic refers to a composition containing at least one live probiotic bacterial strain.
  • Probiotics are live bacteria or yeast that, when consumed, confer a health benefit to the host. Probiotics are said to restore the balance of bacteria in the gut after disruption due to long-term antibiotic use or gastrointestinal disease.
  • compositions of the one or more probiotics may or may not be tailored to address any deficiency in a subject’s gut microbiome or to enhance a subject’s gut microbiome.
  • the probiotic is considered to be off-the-shelf and comprises a standard one or more microbes to enhance therapy of any kind, including therapy to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels.
  • Such a probiotic may be given to a subject without having prior analysis of their gut microbiome.
  • the probiotic may comprise any one or more microbes associated with efficacious therapy to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels.
  • the probiotic composition is tailored to the specific deficiencies of the gut microbiome of the subject. In some cases, such a customized probiotic may or may not comprise all of the microbes that are considered to be deficient in the subject.
  • the subject may be given one or more probiotic compositions, including compositions that comprise one or more microbes that overcome any deficiencies in the subject’s gut microbiome.
  • the probiotic may be given to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels and/or enhance therapy to treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite levels.
  • the probiotic composition comprises a Lactobacillus composition, such as a L. reuteri composition.
  • a Lactobacillus composition such as a L. reuteri composition.
  • Any strain of L. reuteri may be utilized.
  • strains of L. reuteri include at least ATCC PTA 6475 (MM4-1A), ATCC 6A15::pocR, ATCC PTA 4659 (MM2- 3), ATCC PTA 5289 (FJ1), CRL1324, 1048, 1063, 1073, 173.5, 27.4, atcc55739, atcc53608, jw2015, jw2019, ks6, lem83, lr85573, tmwll294, 10c2, p97, pg3b, 32, 676, cp447, 6sl5, cp415, lpl67.67, lpal, tmwll46, tmwll37, 100.93, ad23,
  • the strains of L. reuteri include at least ATCC PTA 6475, DSM17938, ATCC PTA4659, ATCC PTA 5289, ATCC 55730, CRL1324, DSM20016, CF4-6G, ATCC55730, CF48-3A1, M27U15, or a combination thereof. In some embodiments, the strain of L. reuteri includes at least ATCC PTA 6475.
  • the L. reuteri may be obtained from any suitable source, including commercially, from a research laboratory, as a gift, or isolated from nature. Isolation methods for L. reuteri are known in the art, including at least those described in U.S. Patent No. 5,439,678 and U.S. Patent No. 5,849,289 (both incorporated by reference herein in their entirety), for example.
  • L. reuteri is not delivered as a live microorganism, but instead a specific extract or fraction produced from the live microorganism may be equally bioactive. This has been demonstrated, for example, in the case of wound healing in animals, in which consumed sonicated mixtures or lysates prepared from live or dead L. reuteri are biologically active in a wound healing model (Poutahidis et al., 2013; Varian et al., 2016).
  • the mixtures, lysates or extracts may be delivered in pill form or any other standard delivery mode (food or liquid beverages, yogurt, dry powder mixed in water or beverage in which such mixtures can be effectively dosed and consumed in a biologically active format.
  • compositions comprising the bacteria selected using the methods described above are provided.
  • the compositions may include a pharmaceutically acceptable carrier, diluent and/or excipient, in some cases.
  • the composition may include more than one bacterial isolate.
  • the compositions may be formulated for delivery in food, water, via oral gavage, via an aerosol or sprayable product, drops, powder freeze-dried, extract, pill and/or via suppository, in specific cases.
  • the L. reuteri composition is formulated to target a region in the gastrointestinal tract, including any portion of the gastrointestinal tract.
  • the L. reuteri composition is formulated to be delivered to any portion of the gastrointestinal tract.
  • the composition can be formulated as a frozen composition, e.g., flash frozen, dried or lyophilized for storage and/or transport.
  • strains or compositions encompassed herein may be administered in a variety of ways known or available to those skilled in the art.
  • the strains or compositions may be administered in the form of a pharmaceutical, nutraceutical, added to food and/or water, and/or provided in aerosolized or sprayable form for administration by inhalation.
  • the strains and compositions described herein may be provided as liquid suspensions, lyophilized or freeze dried powders or frozen concentrates for addition to target regions other than a subject, as examples.
  • the probiotic microorganisms are preferably not freeze- or spray-dried in a ready-to-use product, because the microorganisms preferably should be as robust as possible when entering the intestines in order to successfully compete with the pro-inflammatory part of the microflora already present in the intestines.
  • some strains may be sufficiently robust and/or some protocols sufficiently gentle to allow spray- or freeze-dried probiotic microorganisms in a ready-to-use product.
  • the probiotic composition is directly or indirectly delivered to the digestive tract of the subject.
  • the probiotic composition is administered to the subject by a route selected from the group consisting of oral, rectal (e.g., by enema), nasal, and via naso/oro-gastric gavage.
  • the probiotic composition is delivered to the subject in a form of a liquid, foam, cream, spray, powder, or gel.
  • the probiotic composition comprises a buffering agent (e.g., sodium bicarbonate, infant formula or sterilized human milk, or other agents which allow bacteria to survive and grow (e.g., survive in the acidic environment of the stomach and to grow in the intestinal environment), along with preservatives, stabilizers, binders, compaction agents, lubricants, dispersion enhancers, disintegration agents, antioxidants, flavoring agents, sweeteners, and coloring agents.
  • a buffering agent e.g., sodium bicarbonate, infant formula or sterilized human milk, or other agents which allow bacteria to survive and grow (e.g., survive in the acidic environment of the stomach and to grow in the intestinal environment)
  • preservatives e.g., sodium bicarbonate, infant formula or sterilized human milk, or other agents which allow bacteria to survive and grow (e.g., survive in the acidic environment of the stomach and to grow in the intestinal environment)
  • preservatives e.g., sodium bicarbonate, infant formula or steriliz
  • the composition can be administered alone or in combination with a carrier, such as a pharmaceutically acceptable carrier or a biocompatible scaffold.
  • a carrier such as a pharmaceutically acceptable carrier or a biocompatible scaffold.
  • formulations of the composition comprise an ingestible carrier, which may be a pharmaceutically acceptable carrier such as a capsule, tablet or powder.
  • the ingestible carrier is a food product such as acidified milk, yogurt (frozen or non-frozen), milk powder, milk concentrate, cheese spreads, dressings and/or beverages.
  • the composition further comprises a protein and/or peptide, in particular proteins and/or peptides that are rich in glutamine/glutamate, a lipid, a carbohydrate, a vitamin, mineral and/or trace element, for example.
  • the formulation of the composition may further comprise an adjuvant, a drug, a biological compound, or a mixture thereof.
  • the formulation of the composition may be a food stuff or a medicament, in certain embodiments.
  • Additional formulations of the composition that are suitable for other modes of administration include oral formulations.
  • Oral formulations include such normally employed excipients such as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. These formulations take the form of solutions, suppositories, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain about 10% to about 95% of active ingredient, preferably about 25% to about 70%.
  • the compositions are formulated for oral administration.
  • Oral administration may be achieved using a chewable formulation, a dissolving formulation, an encapsulated/coated formulation, a multi-layered lozenge (to separate active ingredients and/or active ingredients and excipients), a slow release/timed release formulation, or other suitable formulations known to persons skilled in the art.
  • the word “tablet” is used herein, the formulation may take a variety of physical forms that may commonly be referred to by other terms, such as lozenge, pill, capsule, or the like.
  • compositions of the present disclosure may be formulated for oral administration, other routes of administration can be employed, however, including, but not limited to, subcutaneous, intramuscular, intradermal, transdermal, intraocular, intraperitoneal, mucosal, vaginal, rectal, and intravenous.
  • the disclosed compositions are formulated as a capsule.
  • the capsule may be a hollow, generally cylindrical capsule formed from various substances, such as gelatin, cellulose, carbohydrate or the like.
  • compositions are formulated as a suppository.
  • the suppository may include but is not limited to the bacteria and one or more carriers, such as polyethylene glycol, acacia, acetylated monoglycerides, camuba wax, cellulose acetate phthalate, com starch, dibutyl phthalate, docusate sodium, gelatin, glycerin, iron oxides, kaolin, lactose, magnesium stearate, methyl paraben, pharmaceutical glaze, povidone, propyl paraben, sodium benzoate, sorbitan monoleate, sucrose talc, titanium dioxide, white wax and coloring agents.
  • carriers such as polyethylene glycol, acacia, acetylated monoglycerides, camuba wax, cellulose acetate phthalate, com starch, dibutyl phthalate, docusate sodium, gelatin, glycerin, iron oxides, kaolin, lactose, magnesium stearate,
  • the disclosed compositions are prepared as a tablet.
  • the tablet may include the bacteria and one or more tableting agents, such as dibasic calcium phosphate, stearic acid, croscarmellose, silica, cellulose and cellulose coating.
  • the tablets may be formed using a direct compression process, though those skilled in the art will appreciate that various techniques may be used to form the tablets.
  • the probiotic compositions of the present disclosure may be prepared as a powder that is intended to be dissolved in a liquid, such as water, milk, juice, and yogurt. It is understood that individual liquids may be mixed together where appropriate.
  • a liquid such as water, milk, juice, and yogurt.
  • individual liquids may be mixed together where appropriate.
  • the probiotic formulation may be combined with fruit juice and yogurt or milk and yogurt to make probiotic yogurt shakes.
  • the probiotic formulation may also be combined with milk and ice cream to make probiotic milk shakes. Flavorings for the probiotic liquid formulations contemplated in the disclosure are known to those of ordinary skill in the art.
  • the formulation of the compositions may be provided in food and/or beverage, in specific embodiments.
  • Particular examples include dairy products and/or probiotic-fortified foods.
  • tablets, capsules, gums, or powders containing the bacteria in freeze-dried form are also available.
  • the L. reuteri is provided in a pill or in yogurt, for example.
  • the formulation may be provided in infant formula and/or cereal (infant or otherwise), in some cases.
  • the L. reuteri composition and related methods of the present disclosure utilize particular formulation methods.
  • the carrier may further comprise a disintegrant, a glidant, and/or a lubricant, such as is described in U.S. Patent 9,084,434 (incorporated herein by reference in its entirety), for example, to facilitate having a greater shelf life and/or half-life of the formulation.
  • the disintegrant may be any suitable disintegrant such as, for example, a disintegrant selected from the group consisting of sodium croscarmellose, crospovidone, gellan gum, hydroxypropyl cellulose, starch, and sodium starch glycolate.
  • the glidant may be any suitable glidant such as for example, a glidant selected from the group consisting of silicon dioxide, colloidal silicon dioxide, and talc.
  • the lubricant may be any suitable lubricant such as for example, a lubricant selected from the group consisting of calcium stearate, magnesium stearate, stearic acid, sodium stearyl fumerate, and vegetable based fatty acids.
  • the carrier is present in the composition in a range of approximately 30% w/w to approximately 98% w/w; this weight percentage is a cumulative weight percentage taking into consideration all ingredients present in the carrier.
  • the composition of the present invention may be an oral dosage form, a powder that is mixed into a liquid, or a chewing gum.
  • the oral dosage form may be selected from the group consisting of tablets, caplets, and capsules, wherein the tablets and caplets may be solid or chewable.
  • the composition is a powder, it may be mixed into a liquid that is selected from the group consisting of water, milk, juice, and yogurt.
  • the gum may be soft gum or hard chewing gum tablets.
  • probiotic species such as mannitol, sorbitol, alone or together with the additional sugar alcohol lactitol and/or a phytonutrient, such as oligomeric proanthocyanidins (OPC)
  • sugar alcohols such as mannitol, sorbitol
  • a phytonutrient such as oligomeric proanthocyanidins (OPC)
  • the L. reuteri formulation of the compositions comprises one or more excipients.
  • excipients that may be used to formulate appropriate dosage forms include binders, disintegrants, lubricants, coatings, plasticizers, compression agents, wet granulation agents, and sweeteners, all of which are known to those of ordinary skill in the art to which the invention pertains. All of the following examples are provided by way of illustration and not limitation. Binders are used where appropriate to help the dosage form ingredients still together. Examples of binders include carbopol, povidone, and xanthan gum.
  • Lubricants are generally always used in the manufacture of dosage forms by direct compression in order to prevent the compacted powder mass from sticking to the equipment during the tabletting or encapsulation process.
  • examples of lubricants include calcium stearate, magnesium stearate, stearic acid, sodium stearyl fumerate, and vegetable based fatty acids.
  • Disintegrants aid in the break-up of the compacted mass when placed in a fluid environment.
  • disintegrants include sodium croscarmellose, crospovidone, gellan gum, hydroxypropyl cellulose, starch, and sodium starch glycolate. Coatings are used to control the solubility of the drug.
  • coatings include carrageenan, cellulose acetate phthalate, ethylcellulose, gellan gum, matodextrin, methacrylates, methylcellulose, microcrystalline cellulose, and shellac.
  • Plasticizers are used to control the release rate of the drug from the dosage form. Examples of plasticizers include citrate esters, dibutyl sebacate, diethyl phthalate, polyvinylacetate phthalate, and triacetin.
  • Compression agents include calcium carbonate, dextrose, fructose, guar gum, honey, lactose, maltodextrin, maltose, mannitol, microcrystalline cellulose, molasses, sorbitol, starch, and sucrose.
  • Wet granulation agents include calcium carbonate, lactose, maltodextrin, mannitol, microcrystalline cellulose, povidone, and starch.
  • Sweeteners include aspartame, dextrose, fructose, honey, lactose, maltodextrin, maltose, mannitol, molasses, monoammonium glycyrrhizinate, sorbitol, sucralose, and sucrose.
  • Excipients that are generally used in the manufacture of chewable tablets include by way of illustration and not limitation, dextrose, fructose, guar gum, lactose, maltodextrin, maltose, mannitol, microcrystalline cellulose, and sorbitol. As is evident from the foregoing list, many of the same ingredients may be used for various different purposes in various different dosage forms.
  • Embodiments of the disclosure include one or more formulations of a composition comprising a L. reuteri strain suitable for delivery to a subject in need thereof.
  • a L. reuteri composition may be provided as a formulation to the subject in any suitable form, and in particular embodiments, the bacteria of the formulation are live, although in certain embodiments, the bacteria of the formulation are dead.
  • the probiotic compositions of the disclosure can thus comprise, without limitation, e.g., live bacterial cells, conditionally lethal bacterial cells, inactivated bacterial cells, killed bacterial cells, spores (e.g., germination-competent spores), recombinant carrier strains, cell extract, and bacterially-derived products (natural or synthetic bacterially-derived products such as, e.g., bacterial antigens or bacterial metabolic products).
  • GI gastrointestinal
  • the present disclosure also comprises administering “bacterial analogues”, such as recombinant carrier strains expressing one or more heterologous genes derived from the relevant bacterial species.
  • bacterial analogues such as recombinant carrier strains expressing one or more heterologous genes derived from the relevant bacterial species.
  • the use of such recombinant bacteria may allow the use of lower therapeutic amounts due to higher protein expression.
  • the probiotic composition is reconstituted from a lyophilized preparation.
  • said probiotic composition comprises a buffering agent to adjust pH to a suitable number, such as 7.0.
  • a bacterial composition may comprise a therapeutically effective amount of one or more bacteria.
  • a “therapeutically effective” amount of a bacterium describes an amount sufficient to be effective in treating a desired condition, for example, neutropenic fever.
  • a therapeutically effective amount of isolated or purified populations of bacteria administered to a human will be at least about lxlO 3 colony forming units (CFU) of bacteria or at least about lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 CFU (or any derivable range therein).
  • CFU colony forming units
  • a single dose will contain bacteria (such as a specific bacteria or species, genus, or family described herein) present in an amount of least, at most, or about lxlO 3 , lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 or more CFU (or any derivable range therein).
  • bacteria such as a specific bacteria or species, genus, or family described herein
  • a single dose will contain at least, at most, or about lxlO 3 , lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 or greater than lxlO 15 CFU (or any derivable range therein) of total bacteria.
  • a therapeutically effective amount of each isolated or purified population of bacteria that is administered will be at least about lxlO 3 cells of bacteria or at least about lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 cells (or any derivable range therein).
  • a single dose will contain bacteria (such as a specific bacteria or species, genus, or family described herein) present in an amount of at least, at most, or about lxlO 3 , lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 or more cells (or any derivable range therein).
  • bacteria such as a specific bacteria or species, genus, or family described herein
  • a single dose will contain at least, at most, or about lxlO 3 , lxlO 4 , lxlO 5 , lxlO 6 , lxlO 7 , lxlO 8 , lxlO 9 , lxlO 10 , lxlO 11 , lxlO 12 , lxlO 13 , lxlO 14 , lxlO 15 or greater than lxlO 15 cells (or any derivable range therein) of total bacteria.
  • each dose of the L. reuteri composition may comprise a certain range of bacterial cells.
  • the L. reuteri composition comprises a range of about 10 3 cells to about 10 13 cells.
  • each dose is in the range of about 10 5 cells to about 10 11 cells, about 10 5 to about 10 10 cells, about 10 7 cells to about 10 10 cells, and so forth.
  • each dose is in the range of about 10 cells to about 10 1 cells.
  • composition comprising the desired microbe(s), e.g., L. reuteri
  • the desired microbe(s) can be administered alone or in combination with one or more additional probiotic, prebiotic, neutraceutical, or therapeutic agents (e.g. , BH4 or one or more derivatives thereof).
  • Administration “in combination with” one or more further additional probiotic, neutraceutical, or therapeutic agents includes both simultaneous (at the same time) and consecutive administration in any order. Administration can be chronic or intermittent, as deemed appropriate by the supervising practitioner, including in view of any change in any undesirable side effects.
  • the L. reuteri composition is administered prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the L. reuteri composition thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the L. reuteri composition is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) prior to the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the L. reuteri composition is administered after the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the L. reuteri composition is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4,
  • At least 1, 2, 3, 4, 5, 6, or 7 doses (or any derivable range therein) of the L. reuteri composition thereof is administered at least, at most, or about 1, 2, 3, 5, 6, 12, 24 hours or 2, 3, 4, 6, 8, 10, days or 2, 3, 4, 5, 6, 7, or 8 weeks (or any derivable range therein) after the additional probiotic, prebiotic, neutraceutical, or therapeutic agents.
  • the formulation comprising a L. reuteri strain suitable for delivery to a subject in need thereof further comprises vitamins and/or fish oil, for example.
  • the formulation comprising a L. reuteri strain suitable for delivery to a subject in need thereof further comprises a prebiotic material, which is a food ingredient that promotes the growth of the L. reuteri , such as that stimulate the survival, growth or activity of L. reuteri or promote its persistence in the gastrointestinal tract.
  • prebiotic refers to a selectively fermented ingredient that induces specific changes to the composition and/or activity of gastrointestinal microflora to confer benefits upon host well-being and health.
  • prebiotics include but are not limited to simple sugars, complex carbohydrates, fibers, fats, proteins or natural products, such as vitamins, minerals, polyphynols, inulin, arabinoxylan, xylose, soluble fiber dextran, soluble corn fiber, polydextrose, lactose, N-acetyl-lactosamine, glucose, galactose, fructose, rhamnose, mannose, uronic acids, 3’-fucosyllactose, 3’-sialylactose, 6’-sialyllactose, lacto-N-neotetraose, 2’-2’-fucosyllactose, trans-galactooligosaccharides, glucooligosaccharides, isomaltooligosaccharides, lactosucrose, polydextrose, soybean oligosaccharides, and arabinose, cellobiose, fructose, fucos
  • the prebiotic material promotes or stimulates the survival, growth, or activity of L. reuteri or promotes the persistence of L. reuteri in the gastrointestinal tract.
  • An effective amount of L. reuteri may be provided to a subject in need thereof and may or may not be provided with one or more additional probiotics, such as one or more additional probiotic bacteria and/or yeast.
  • the additional probiotic bacteria and/or yeast may be in the same composition as the L. reuteri , or the additional probiotic bacteria and/or yeast may be in a different composition as the L. reuteri.
  • the additional probiotic bacteria and/or yeast may or may not be given to the subject at the same time as the L. reuteri.
  • the additional probiotic bacteria and/or yeast may assist in treating and/or preventing at least a social behavioral deficiency, oxytocinergic system dysfunction, or abnormal biopterin metabolite level and/or the additional probiotic bacteria may be useful for treating and/or preventing at least one symptom of another medical condition.
  • Other medical conditions that may be treated and/or prevented at the same time with other probiotics include at least anxiety, cognition, schizophrenia, bipolar disorder, mood disorder, seasonal affective disorder, irritable bowel syndrome, inflammatory bowel disease (IBD), infectious diarrhea (caused by viruses, bacteria, or parasites), antibiotic -related diarrhea, skin conditions (such as eczema), urinary and vaginal conditions, allergies and colds, and oral conditions.
  • the additional probiotic bacteria and/or yeast may be of any kind, so long as it is not pathogenic in the amount provided to the subject.
  • the additional probiotic bacteria and/or yeast may be Lactobacillus acidophilus, Lactobacillus bifidus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus salivarius, Bacteroides uniformis, Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium animalis subsp.
  • lactis Bifidobacterium longum, Bifidobacterium pseudolongum, Collinsella unclassified, Enterococcus faecalis, Enterococcus faecium, Helicobacter hepaticus, Olsenella unclassified, Parabacteroides distasonis, Streptococcus thermophilis, Saccharomyces boulardii, or a combination thereof.
  • one or more other bacteria and/or yeast may be provided in a particular ratio.
  • such a ratio may be 1:1, 1:2, 1:5, 1:10, 1:25, 1:50, and so forth.
  • the particular ratio may be 1:1:1; 1:2:1; 1:10:1; 1:2:2; 1:10:10; 1:100:1; 1:100:100, and so forth.
  • compositions are administered in a manner compatible with the dosage formulation, and in such amount as will be therapeutically or prophylactically effective for the subject being treated.
  • Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and are peculiar to each individual. Suitable regimes for initial administration and boosters are also variable, but are typified by an initial administration followed by subsequent administrations. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability and toxicity of the particular therapeutic substance or other therapies a subject may be undergoing.
  • compositions will be pharmaceutically acceptable or pharmacologically acceptable.
  • pharmaceutically acceptable or “pharmacologically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal, or human.
  • pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in immunogenic and therapeutic compositions is contemplated.
  • Methods of the disclosure allow for the treatment or prevention a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, by administering a therapeutically effective amount of a formulation comprising L. reuteri and/or BH4 or one or more derivatives thereof.
  • a subject that is the subject for methods and compositions of the disclosure has a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the subject has a neurodevelopmental disorder, a psychiatric disorder, a neurodegenerative disorder, a cardiovascular disorder, and/or a metabolic disorder, and such a disorder may have any cause, including deprivation, genetic and metabolic diseases, immune disorders, infectious diseases, nutritional factors, physical trauma, and toxic and environmental factors, for example; the disorder may have two or more causes.
  • a treatment regimen is for a subject with a neurodevelopmental disorder having as a symptom a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with a psychiatric disorder having as a symptom a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with a neurodegenerative disorder having as a symptom a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with a cardiovascular disorder having as a symptom a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with a metabolic disorder having as a symptom a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with ASD having as a symptom a social behavior deficiency.
  • a treatment regimen is for a subject with William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease having as a symptom an oxytocinergic system dysfunction.
  • a treatment regimen is for a subject with a cardiovascular disorder, for example, cardiovascular disease, having as a symptom abnormal biopterin metabolite levels.
  • a treatment regimen is for a subject with a metabolic disorder, or example, BH4-deficient hyperphenylalaninemia and/or tyrosinemia, having as a symptom abnormal biopterin metabolite levels.
  • a subject having a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels is provided an effective amount of a formulation comprising at least an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof to treat or prevent a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the term “therapeutically effective amount” is synonymous with “effective amount,” “therapeutically effective dose,” and/or “effective dose,” and refers to an amount of an agent sufficient to produce a desired result or exert a desired influence on the particular condition being treated.
  • a therapeutically effective amount is an amount sufficient to ameliorate at least one symptom, behavior or event, associated with a pathological, abnormal or otherwise undesirable condition, or an amount sufficient to prevent or lessen the probability that such a condition will occur or re-occur, or an amount sufficient to delay worsening of such a condition.
  • the effective amount refers to the amount of a formulation comprising L. reuteri and/or BH4 or one or more derivatives thereof that can treat or prevent a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels in a subject.
  • the effective amount may vary depending on the organism or individual treated.
  • treatment refers to intervention in an attempt to alter the natural course of the subject being treated, and may be performed either for prophylaxis or during the course of pathology of a disease or condition. Treatment may serve to accomplish one or more of various desired outcomes, including, for example, preventing occurrence or recurrence of disease, alleviation or reduction in severity of symptoms, and diminishment of any direct or indirect pathological consequences of the disease, preventing disease spread, lowering the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.
  • a regimen having a first phase for an initial treatment, which may last for days, weeks, months, or years, and then another phase for maintenance, which may last for weeks, months, or years.
  • Such a treatment may be administered one or more times a day, including about 1, 2, 3, or more times per day, for a period sufficient to stabilize the gut flora.
  • a maintenance phase a subject is provided a lesser amount of the bacteria and/or fewer administrations than the initial treatment phase.
  • the duration of a treatment regimen may be dependent on each individual patient and the stage of the medical condition.
  • a continued treatment for a certain period of time occurs until a detectable improvement in a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the improved social behavioral deficiency, oxytocinergic system dysfunction, and/or BH4 levels is maintained by additional treatment, although the additional treatment may be reduced in frequency and/or volume.
  • an initial treatment regimen comprising an effective amount of a formulation comprising at least an effective amount of L. reuteri may comprise of at least about 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , or 10 10 cells per dose or any range or value derivable therein.
  • an initial treatment regimen comprising an effective amount of a formulation comprising at least an effective amount of BH4 or one or more derivatives thereof may comprise at least about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, and 200, 300, 400, 500, 1000 pg/kg, mg/kg, pg/day, or mg/day or any range or value derivable therein.
  • an initial treatment regimen comprises an effective amount of a formulation comprising at least an effective amount of L. reuteri , which may comprise of at least about 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , or 10 10 cells per dose or any range or value derivable therein, and at least an effective amount of BH4 or one or more derivatives thereof, which may comprise at least about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, and 200, 300, 400, 500, 1000 pg/kg, mg/kg, pg/day, or mg/day or any range or value derivable therein.
  • abnormal levels of BH4 refers to BH4 levels that are greater than or less than BH4 levels in a standard or control reference.
  • the standard or control reference may comprise the BH4 levels in a healthy individual, for example.
  • the subject may have or may be at risk of having one or more of BH4-deficient hyperphenylalaninemia and/or tyrosinemia.
  • the subject may have or may be at risk of having cardiovascular disease.
  • the subject may have or may be at risk of having a neurodevelopment disorder, autism spectrum disorder, or other neurological disorder associated with social dysfunction.
  • the subject may have or may be at risk of having one or more of William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease associated with oxytocinergic system dysfunction.
  • BH4 tetrahydrobiopterin
  • the subject may have impaired social behavior of any kind.
  • the social behavioral deficiency may comprise impaired sociability, preference for social novelty, difficulty in social use of verbal and nonverbal communication, or a combination thereof.
  • the subject may have or may be at risk of having a neurodevelopment disorder, autism spectrum disorder, or other neurological disorder associated with social dysfunction.
  • One of skill in the art may measure one or more social behavior deficiencies in humans in a variety of ways. In specific embodiments, the skilled artisan would employ the standards set forth in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5):
  • the symptoms are not attributable to another medical or neurological condition or to low abilities in the domains or word structure and grammar, and are not better explained by autism spectrum disorder, intellectual disability (intellectual developmental disorder), global developmental delay, or another mental disorder.
  • a subject may be at risk of having at least one social behavior deficiency because the subject has a decreased abundance of L. reuteri in the gut microbiome and/or decreased levels of BH4 and/or one or more derivatives thereof in the gut.
  • a subject may be at risk of having at least one social behavior deficiency because their biological mother was obese, overweight, and/or on a high fat diet, and the identification of such a risk is a part of some methods.
  • risk factors for a social behavior deficiency are environmental (diet, pathogen infection, exposure to toxin, pesticides, etc.) or genetic factors (mutations in genes associated with ASD and other neurological disorders), patemal/maternal age and prematurity, or any combination of these factors.
  • the subject was bom from a mother that is obese, overweight, and/or is on a high-fat diet, undergoes immune activation during pregnancy, although in some cases the subject was born from a mother that was not obese, not overweight, and/or not on a high- fat diet.
  • the subject mother or parent
  • the mother or individual exhibit an altered gut microbiota or show alteration in oxytocin levels.
  • BH4 neurodegenerative, psychiatric, cardiovascular, and/or metabolic disorders
  • the subject may have, for example, social behavior deficits, social cognitive/memory deficits, addictive behavior, anxiety/depressive behavior, abnormal feeding behavior, or a combination thereof.
  • the subject may have or may be at risk of having one or more of William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease associated with oxytocinergic system dysfunction.
  • One of skill in the art may measure one or more oxytocinergic system dysfunctions in humans in a variety of ways.
  • immuno-assays such as enzyme linked immunosorbent assay (ELISA) or radioimmunoassay (RIA) or other methods such as chromatography or mass spectrometry.
  • the present disclosure concerns neurodevelopmental and psychiatric disorders including medical conditions in which the oxytocinergic system is dysfunctional, such as William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease, for example.
  • medical conditions in which the oxytocinergic system is dysfunctional such as William Syndrome, Prader-Willi Syndrome, Fragile X syndrome, one or more addictive behaviors, sleep disorder, bipolar disorder, schizophrenia, depression, postpartum depression, anxiety, dystonia, Parkinson’s disease, social cognitive impairment, and/or Alzheimer’s disease, for example.
  • targeting the oxytocin system via administration of L. reuteri and/or BH4 or one or more derivatives thereof is useful to treat addictive behaviors including eating/feeding disorders and drug addiction, sleep disorder, bipolar disorder, and Schizophrenia, for example (Cochran, etal., 2013; MacDonald, etal., 2014; McQuaid, etal., 2014; Samyai, et al, 2014; Kirsch, et al, 2015; Perez-Rodriguez, et al., 2015; Rice, et al, 2015; Romano, et al., 2015; Zik, et al., 2015; Brambilla, et al., 2016; Bukovskaya, et al., 2016; Lee, et al., 2016; Quintana, etal., 2017).
  • the addictive behavior can concern, for example, food, alcohol, drugs, gambling, inhalants, and tobacco.
  • an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof is provided to a subject with or at risk for depression.
  • Depression is one of the most common psychiatric disorders. It causes severe symptoms that affect how one feels, thinks, and handles daily activities, such as sleeping, eating, or working.
  • L. reuteri and/or BH4 or one or more derivatives thereof can promote the production of oxytocin
  • L. reuteri and/or BH4 or one or more derivatives thereof in particular embodiments is useful in the treatment of major depressive disorders.
  • L. reuteri and/or BH4 or one or more derivatives thereof can promote the activity of dopaminergic neurons in a key reward area.
  • reward dopaminergic pathways are altered.
  • reuteri is useful for the treatment of conditions in which the dopaminergic and reward system are usurped, including drug addiction (Hyman, et al., 2006), obesity and food intake (Kenny, et al., 2011; Blum, et al., 2014), dystonia, Parkinson’s disease, and other addictive behaviors, such as gambling (Fenu, et al., 2009; Matsumoto, et al., 2015), for example.
  • the present disclosure improves social cognitive function, such as with neurodegenerative disorders.
  • Social cognitive impairment which can lead to poor quality of life, mental health problems and loneliness (Kennedy, et al., 2012), can be prominent after acute brain damage (for example), such as traumatic brain injury or stroke, and in specific embodiments represents a core feature of the early stages of some chronic neurological disorders, such as frontotemporal dementia, Alzheimer’s disease and Parkinson’s disease (Beer, et al., 2006; Henry, et al., 2016).
  • oxytocin and dopamine are crucial for the formation of social memory (Ferguson, et al., 2000; Gonzalez-Burgos, et al., 2008; Shamay-Tsoory, et al., 2016). Because L.
  • reuteri and/or BH4 or one or more derivatives thereof can promote oxytocin production and dopamine-mediated synaptic plasticity, two systems that are hijacked in disorders exhibiting social cognition problems (Rudelli, et al., 1984; Gibb, et al., 1989; Braak, et al., 1990; Selden, et al., 1994; Bums, et al., 2005; Ross, et al., 2009; Borroni, et al., 2010; Finger, et al., 2011; Tampi, et al., 2017), the formulations disclosed herein are useful for the treatment of social memory and cognitive function in neurological and neurodegenerative disorders.
  • a subject may be at risk of having at least one oxytocinergic system dysfunction because the subject has a decreased abundance of L. reuteri in the gut microbiome and/or decreased levels of BH4 and/or one or more derivatives thereof in the gut.
  • a subject may be at risk of having at least one oxytocinergic system dysfunction because their biological mother was obese, overweight, and/or on a high fat diet.
  • Other risk factors include but are not limited to the presence of gene variants related to nervous system function, for example, Shank3B and Cntnap2, and/or exposure to environmental toxins including but not limited to valproic acid (VPA) during development.
  • the identification of such a risk of having at least one oxytocinergic system dysfunction is a part of some methods.
  • a subject in need thereof is provided an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof for the purpose of improving a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a subject is intentionally provided an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof for the purpose of improving at least one social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite level.
  • L. reuteri and/or BH4 or one or more derivatives thereof may be administered to increase social bonding, improve mood, reduce depression and increase wellbeing in individuals, including friends, family members, colleagues, and partners.
  • At least one symptom of at least one neurodevelopmental disorder is treated with an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof.
  • a subject may be treated for social deficits, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, as well as changes in gut microbiome, by providing an effective amount of a formulation comprising L. reuteri and/or BH4 or one or more derivatives thereof.
  • the gut microbiome of a subject is analyzed or measured or determined or evaluated for the overall diversity of its microbes, irrespective of which microbes are actually present and/or absent.
  • the overall diversity and/or activity of the gut microbiome may be measured in any suitable manner, including for example, gene marker analysis (such as with next generation sequencing), shotgun metagenomics (such as with next generation sequencing), metabolomics (such as with liquid chromatography/gas chromatography-mass spectrometry), metaproteomics (such as with liquid chromatography/gas chromatography- mass spectrometry), and/or metatransciptomics (such as with next generation sequencing).
  • a subject having a gut microbiome having a decreased abundance of L are examples of the gut microbiome having a decreased abundance of L.
  • reuteri has an increased chance of developing a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • a subject having decreased levels of BH4 and/or one or more derivatives thereof has an increased chance of developing a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • Methods of the disclosure include methods of treating or preventing a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, where risk of developing a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, is increased.
  • the subject is at a higher risk than an average person in the general population.
  • the medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels poses a greater risk to the health or life of the subject than such a condition would pose to an average person in the general population.
  • the method is employed for a subject where it is uncertain whether or not risk of developing a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, is increased, whereas in other cases the method is employed for a subject where it is known that the risk of developing a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, is increased.
  • the disclosure encompasses methods and compositions for modulating the gut microbiome activity and/or composition of a subject to treat or prevent a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the modulation may or may not be as a result of analysis of the gut microbiome activity and/or composition prior to or after diagnosing the subject with a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the modulation is a result of analysis of the gut microbiome prior to diagnosing the subject with a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, and the outcome of the analysis determines the nature of the resultant modulation of the gut microbiome.
  • the modulation may comprise providing a therapeutically effective amount of L. reuteri and/or BH4 or one or more derivatives thereof.
  • the subject was determined to have a decreased abundance of L. reuteri in the gut microbiome compared to a control or reference sample, and the modulation may comprise providing an effective amount of L. reuteri to increase the abundance of L. reuteri in the gut of the subject.
  • the subject was determined to have decreased levels of BH4 or one or more derivatives thereof compared to a control or reference sample, and the modulation may comprise providing an effective amount of BH4 or one or more derivatives thereof to increase levels of BH4 or one or more derivatives thereof in the gut of the subject.
  • the control or reference sample is a sample from a healthy subject lacking a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the subject does not exhibit a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels when the formulation comprising an effective amount of L. reuteri and/or BH4 or one or more derivatives thereof is administered.
  • the subject has been diagnosed with a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels.
  • the formulation comprising an effective amount of L.
  • reuteri and/or BH4 or one or more derivatives thereof is administered after the subject has been diagnosed with a medical condition in which at least one symptom is a social behavioral deficiency, oxytocinergic system dysfunction, and/or abnormal biopterin metabolite levels, and the composition may be administered to the subject until the subject is no longer symptomatic.
  • the abundance of L. reuteri and/or the levels of BH4 or one or more derivatives thereof in the gut of a subject were determined from a fecal sample from the subject.
  • the effective amount of the formulation comprising L. reuteri and/or BH4 or one or more derivatives thereof indirectly or directly improves social interaction and/or the oxytocinergic system dysfunction and/or BH4 levels because it indirectly or directly induces synaptic potentiation (LTP) in the ventral tegmental area (VTA) of the subject.
  • the effective amount of the comprising L. reuteri and/or BH4 or one or more derivatives thereof indirectly or directly increases the number of oxytocin immunoreactive neurons in the hypothalamus.
  • Embodiments of the disclosure impact the consequences of diet, gut microbiota dysbiosis, VTA plasticity, abnormal social behavior, oxytocinergic system dysfunction, and BH4 levels by providing an effective L. reuteri and/or BH4-based therapy for a subject.
  • a subject is provided a therapy or preventative for one or more social behavior deficiencies and/or oxytocinergic system dysfunctions and/or abnormal biopterin metabolite levels in addition to the compositions encompassed herein.
  • they may be provided the following in addition to and/or in combination with formations comprising L. reuteri and/or BH4 or one or more derivatives thereof: a) other bacteria species, b) prebiotics; c) other nutrients (e.g., vitamins, lipids, proteins, etc.); and d) behavioral therapy.
  • the analysis of the gut microbiome to determine its content may be performed by any suitable method.
  • the analysis may begin with collection of a suitable sample, such as stool, tissue biopsy, or a combination thereof.
  • a suitable sample such as stool, tissue biopsy, or a combination thereof.
  • one may collect the whole stool, homogenize it immediately (e.g., with a blender or a tissue homogenizer), then flash freeze the homogenate in liquid nitrogen or in dry ice/ethanol slurry, with an aliquot preserved in a certain percentage of glycerol in suitable media for culturing.
  • the subject that obtains the sample may or may not be the subject that performs the analysis.
  • the sample is stored prior to analysis, whereas in other cases the sample is analyzed without storage.
  • the gut microbiome is analyzed based on shotgun sequencing of nucleic acid of the microbe(s), including shotgun metagenomics sequencing, such as to provide more in-depth reads.
  • the majority or substantially all of the genomic DNA for a microbe is analyzed instead of a specific region of DNA.
  • analysis of a specific region of DNA is utilized, such as with 16S rRNA sequencing.
  • Other analysis methods may be utilized, either alone or with other methods.
  • culturing may be utilized as a detection method.
  • Assay panels that target a set of known microbes or genes thereof may be utilized.
  • Stool samples may be processed through nucleic acid extraction followed by complementary DNA synthesis and subsequent amplification using mixtures of primers specific for a given range of organisms.
  • Either genomic DNA or PCR product may then be qualified and quantified, such as through a hybridization array using a fluorescence-based measure or a melt curve analysis.
  • quantitative PCR and reverse-transcription quantitative PCR may be utilized.
  • amplicon analyses are employed in which a specific region of DNA is amplified by orders of magnitude using various methods including PCR.
  • the PCR primers match a specific region, such as the 16S rRNA for bacteria.
  • Bacterial 16S rRNA genes contain 9 hypervariable regions (V1-V9) that show sequence diversity and can be used as a barcode-like method to differentiate many bacterial taxa, including at the species level.
  • next-generation sequencing may be performed to read the sequences.
  • shotgun metagenomics is utilized that fragments all the DNA from a sample into small pieces, sequences these fragments, and then the sequenced fragments are arranged accordingly to provide information on a grander scale for the microbe identification.
  • methods involve obtaining a sample from a subject.
  • the methods of obtaining provided herein may include methods of biopsy such as fine needle aspiration, core needle biopsy, vacuum assisted biopsy, incisional biopsy, excisional biopsy, punch biopsy, shave biopsy, or skin biopsy.
  • the sample may be obtained from any of the tissues provided herein that include but are not limited to non-cancerous or cancerous tissue and non- cancerous or cancerous tissue from the serum, gall bladder, mucosal, skin, heart, lung, breast, pancreas, blood, liver, muscle, kidney, smooth muscle, bladder, colon, intestine, brain, prostate, esophagus, or thyroid tissue.
  • the sample may be obtained from any other source including but not limited to blood, sweat, hair follicle, buccal tissue, tears, menses, feces, or saliva.
  • any medical professional such as a doctor, nurse or medical technician may obtain a biological sample for testing.
  • the biological sample can be obtained without the assistance of a medical professional.
  • a sample may include but is not limited to, tissue, cells, or biological material from cells or derived from cells of a subject.
  • the biological sample may be a heterogeneous or homogeneous population of cells or tissues.
  • the biological sample may be obtained using any method known to the art that can provide a sample suitable for the analytical methods described herein.
  • the sample may be obtained by non-invasive methods including but not limited to: scraping of the skin or cervix, swabbing of the cheek, saliva collection, urine collection, feces collection, collection of menses, tears, or semen.
  • the sample may be obtained by methods known in the art.
  • the samples are obtained by biopsy.
  • the sample is obtained by swabbing, endoscopy, scraping, phlebotomy, or any other methods known in the art.
  • the sample may be obtained, stored, or transported using components of a kit of the present methods.
  • multiple samples such as multiple esophageal samples may be obtained for diagnosis by the methods described herein.
  • multiple samples such as one or more samples from one tissue type (for example esophagus) and one or more samples from another specimen (for example serum) may be obtained for diagnosis by the methods.
  • samples such as one or more samples from one tissue type (e.g . esophagus) and one or more samples from another specimen (e.g. serum) may be obtained at the same or different times. Samples may be obtained at different times are stored and/or analyzed by different methods. For example, a sample may be obtained and analyzed by routine staining methods or any other cytological analysis methods.
  • tissue type e.g . esophagus
  • serum e.g. serum
  • the biological sample may be obtained by a physician, nurse, or other medical professional such as a medical technician, endocrinologist, cytologist, phlebotomist, radiologist, or a pulmonologist.
  • the medical professional may indicate the appropriate test or assay to perform on the sample.
  • a molecular profiling business may consult on which assays or tests are most appropriately indicated.
  • the patient or subject may obtain a biological sample for testing without the assistance of a medical professional, such as obtaining a whole blood sample, a urine sample, a fecal sample, a buccal sample, or a saliva sample.
  • the sample is obtained by an invasive procedure including but not limited to: biopsy, needle aspiration, endoscopy, or phlebotomy.
  • the method of needle aspiration may further include fine needle aspiration, core needle biopsy, vacuum assisted biopsy, or large core biopsy.
  • multiple samples may be obtained by the methods herein to ensure a sufficient amount of biological material.
  • the sample is a fine needle aspirate of an esophageal or a suspected esophageal tumor or neoplasm.
  • the fine needle aspirate sampling procedure may be guided by the use of an ultrasound, X-ray, or other imaging device.
  • the biological sample may be obtained from a subject directly, from a medical professional, from a third party, or from a kit provided by a third party.
  • the subject, a medical professional, or a third party may be provided with suitable containers and excipients for storage and transport of the biological sample.
  • a medical professional need not be involved in the initial sample acquisition.
  • a subject may alternatively obtain a sample through the use of an over the counter (OTC) kit.
  • OTC kit may contain a means for obtaining said sample as described herein, a means for storing said sample for inspection, and instructions for proper use of the kit.
  • a sample suitable for use may be any material containing tissues, cells, nucleic acids, genes, gene fragments, expression products, gene expression products, or gene expression product fragments of a subject to be tested. Methods for determining sample suitability and/or adequacy are known in the art.
  • the subject may be referred to a specialist such as an oncologist, surgeon, or endocrinologist.
  • the specialist may likewise obtain a biological sample for testing or refer the subject to a testing center or laboratory for submission of the biological sample.
  • the medical professional may refer the subject to a testing center or laboratory for submission of the biological sample.
  • the subject may provide the sample.
  • a molecular profiling business may obtain the sample.
  • Kits are also included as part of the disclosure. Kits for implementing methods of the invention described herein are specifically contemplated. In some embodiments, there are kits for treating social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels in a subject and/or preventing onset of at least one symptom of social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels in a subject. The kit may be provided for treatment and/or prevention of social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels in a subject having an altered gut microbiome.
  • the container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe or other container means, into which a Lactobacillis component (including L. reuteri ) and/or a metabolite component (including BH4 or one or more derivatives thereof) may be placed, and preferably, suitably aliquoted.
  • a Lactobacillis component including L. reuteri
  • a metabolite component including BH4 or one or more derivatives thereof
  • the kit also will generally contain a second, third or other additional container into which the additional components (such as another bacteria or metabolite) may be separately placed.
  • the kits of the present disclosure also will typically include a means for containing the compositions, and any other reagent containers in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained.
  • the liquid solution is an aqueous solution, with a sterile aqueous solution being particularly preferred.
  • the components of the kit may be provided as a stab or as a dried powder(s).
  • the powder can be reconstituted by the addition of a suitable solvent. It is envisioned that the solvent may also be provided in another container means.
  • the components of the kits may be packaged either in aqueous media or in lyophilized form.
  • kits will also include instructions for employing the kit components as well the use of any other reagent not included in the kit. Instructions may include variations that can be implemented.
  • kits for treating social behavioral deficiencies, oxytocinergic system dysfunctions, and/or abnormal biopterin metabolite levels.
  • Other treatments can involve behavioral treatments, medicines, or both.
  • the subject may have additional medical conditions such as sleep disturbance, seizures, and/or gastrointestinal (GI) distress, and the kits may also have therapeutic compositions to address these conditions.
  • GI gastrointestinal
  • Cntnap2 To study the role of host microbiome interactions in complex behaviors, the inventors employed the Cntnap2 / mouse model for neurodevelopmental disorders. Consistent with some of the behavioral abnormalities observed in individuals with mutations in CNTNAP2, loss of Cntnap2 [contactin-associated protein-like 2; which encodes the protein Caspr2 (Poliak el al., 1999; Poliak et al., 2001)] in mice leads to social deficits and hyperactivity (Bakkaloglu el al., 2008; O’Roak el al., 2011; Penagarikano et al., 2011; Penagarikano et al., 2015).
  • the inventors first tested social behavior in wild-type (WT; Cntnap2 +/+ ) and knockout (KO; Cntnap2 / ) mice bom from isolated homozygous breeding lines (Isolated lines: WT-I and KO-I, respectively; FIG. 1A, FIG. IB) using the 3-chamber test for sociability and social novelty (FIG. 2A).
  • WT-I mice spent significantly more time interacting with the stranger mouse (Mouse 1) than with the empty cup (Empty), indicating normal sociability (FIG. 1C).
  • KO-I mice exhibited impaired sociability as evidenced by their lack of social preference (FIG.
  • KO-I mice did not prefer to interact with a novel mouse (Mouse 2) over a familiar mouse (Mouse 1) in the social novelty test (FIG. ID). Accordingly, in a reciprocal social interaction test (FIG. 2B), KO-I mice spent less time interacting with each other compared to WT-I mice (FIG. IE). KO-I mice displayed no deficits in olfaction compared to WT-I mice as measured in the buried food test (FIG. 3), indicating that their social deficits were not likely due to impaired olfaction.
  • KO-I mice were hyperactive compared to WT-I mice (FIG. IF, FIG. 1G), consistent with previous reports (Penagarikano el al., 2011; Penagarikano el al., 2015).
  • Cntnap2 A mice generated from isolated breeding (KO- I) show impaired social behavior and hyperactivity.
  • KO-I mice show alterations in their gut microbial ecology using 16S ribosomal RNA (rRNA) gene sequencing.
  • Analysis of beta (between subject) diversity revealed that KO-I mice harbor an intestinal microbial composition distinct from WT-I mice.
  • the inventors used three independent metrics: principal components analysis (PCA) with PhILR (Phylogenetic Isometric Log-Ratio)-transformed Euclidean distances (Silverman et al., 2017) (FIG. 1H) and principal coordinates analysis (PCoA) with Bray-Curtis (FIG. 4A) or unweighted UniFrac dissimilarities (FIG. 4B).
  • PCA principal components analysis
  • PhILR Physical Isometric Log-Ratio
  • KO- I mice have increased diversity compared to WT-I mice (FIG. 4C, FIG. 4D).
  • 33 amplicon sequence variants (AS Vs) were differentially abundant in KO-I mice compared to WT- I mice (FIG. II, FIG. 4E) as well as 33 predicted microbial metabolic pathways inferred by PICRUSt2 (FIG. 4F, FIG. 4G).
  • Cntnap2 mice originating from the isolated line (KO-I) show differences in their gut microbiota.
  • the oxytocinergic system modulates numerous aspects of social behaviors and is implicated in ASD (Donaldson and Young, 2008; LoParo and Waldman, 2015) and has been reported to be deficient in Cntnap2 A mice (Penagarikano el al., 2015).
  • Cntnap2 A mice the paraventricular nuclei (PVN) of the hypothalamus, where oxytocin is primarily synthesized, contain less oxytocin producing neurons and oxytocin treatment reverses the social deficits in the mutant mice (Penagarikano et al., 2015). Accordingly, the inventors found that intranasal oxytocin treatment reversed the social deficits, but not the hyperactivity, in Cntnap2 A (KO-I) mice (FIG. 5).
  • Lactobacillus reuteri promotes oxytocin levels in plasma and the PVN (Buffington et al., 2016; Poutahidis el al., 2013; Sgritta et al., 2019) and reverses social deficits in several mouse models for ASD (Buffington el al., 2016; Sgritta et al., 2019; Tabouy l al., 2018).
  • the same manipulation failed to do so in mice with deficient oxytocin receptors (Sgritta et al., 2019).
  • VTA ventral tegmental area
  • VTA Brain regions responding to naturally rewarding stimuli, including the VTA, are crucially involved in social behavior. More specifically, oxytocin-expressing neurons in the PVN project to dopaminergic (DA) neurons of the VTA (Melis et al., 2007) and the activation of DA neurons by oxytocin within this circuit is critically required for social behavior (Gunaydin el al., 2014; Hung et al., 2017; Xiao et al., 2017).
  • DA dopaminergic
  • mEPSCs miniature excitatory post synaptic currents
  • mPFC medial prefrontal complex
  • FIG. 10 the amplitude and frequency of mEPSCs in pyramidal neurons from mPFC were reduced in KO-I mice compared to WT-I (FIG. 10).
  • Treatment with L. reuteri failed to reverse the changes in mEPSCs in KO-I (FIG. 10).
  • BH4 is the biologically active biopterin molecule that serves as co-factor for a number of important enzymes that produce neuroactive molecules such as dopamine, serotonin, and nitric oxide (Thony et al., 2000). Because L. reuteri promotes the level of metabolites in the BH4 pathway (FIG. 11C, FIG. 11D) and reverses the social deficits but not the hyperactivity phenotype in KO-I mice (FIGs. 8B-E, FIG. 8G), and because the social deficits in KO-I mice are mediated by the microbiome (FIG. 1), the inventors next examined whether BH4 treatment alone was sufficient to selectively reverse the social deficits in KO-I mice.
  • KO-I mice were treated with BH4 (or vehicle) via oral gavage (FIG. HE).
  • treatment with BH4 alone reversed the social deficits (FIG. 11F, FIG. 11G, FIG. Ill) and the impaired synaptic plasticity in VTA DA neurons after social interaction (FIG. 11H, FIG. 11J).
  • treatment with BH4 failed to reverse the hyperactivity phenotype in KO-I mice (FIG. 11K, FIG. 11L), further strengthening the notion that the social deficits are mediated by changes in the microbiome and the hyperactivity phenotype is modulated by host genes.
  • L. reuteri reverses social and synaptic plasticity deficits, at least, in part, by modulating the host’s endogenous levels of tetrahydrobiopterin in the gut.
  • L. reuteri signals to the brain and modulates social behavior via the vagus nerve and the oxytocinergic system (Sgritta el al., 2019). More specifically, either surgical cutting of the vagus nerve or inhibition of oxytocin receptors in DA neurons prevents the prosocial effect mediated by L. reuteri (Sgritta et al., 2019).
  • BH4 has been shown to increase oxytocin release in rats (Ciosek and Guzek, 1992; Ciosek et al., 1992) and the inventors found that BH4 failed to reverse social deficits in KO-I mice that were treated with a selective and potent oxytocin receptor antagonist compared to vehicle treated KO-I mice (FIG. 15).
  • mice [0186] Mouse Husbandry.
  • WT C57BL/6J (stock #000664) and Cntnap2 _/ (Cntnap2 mlPele , stock # 017482) (Poliak et al., 2003) were obtained from Jackson Laboratories (Bar Harbor, ME).
  • WT-I and KO-I mice were generated by breeding WT and Cntnap2 / obtained from Jackson Laboratories.
  • Germ- free (GF) mice (C57BL/6J) were housed in a flexible isolator fed with HEPA- filtered air and provided with irradiated food and water at the Baylor College of Medicine gnotobiotic facility. Subject mice were separated by sex at weaning and housed at 2-4 mice per cage.
  • mice both male and female mice were used in this study. Animal care and experimental procedures were approved by Baylor College of Medicine’s Institutional Animal Care and Use Committee in accordance with all guidelines set forth by the U.S. National Institutes of Health. [0187] Behavior Experiments. All behavior experiments were conducted on mice aged 8-12 weeks unless otherwise stated. Mice were habituated to the experimenter for 3 days prior to the start of the behavioral experiment. All experiments were conducted during the light cycle.
  • mice were first habituated for a 10-minute period in an empty 60 x 40 x 23 cm Plexiglas Arena divided into three equally-sized interconnected chambers (left, center, right). During habituation, the subject’s position was continuously tracked using the automated tracking software, AnyMaze. Distance traveled and speed were automatically scored and used for measuring activity levels. Sociability was evaluated during a second 10-minute period in which the subject could interact either with an empty wire cup (Empty) or a wire cup containing a stranger mouse (Mouse 1).
  • Stranger mice were age- and sex-matched to the subject mouse.
  • the interaction time was scored by measuring the time the subject mouse spent sniffing or climbing upon either the empty cup or the cup containing the stranger mouse.
  • the position of the empty cup/stranger mouse in the left or right chamber during the sociability period was counterbalanced between trials, in order to avoid bias.
  • Preference for social novelty was evaluated, in a third 10-minute period, by introducing a second stranger mouse (Mouse 2) into the previously empty wire cup.
  • the time spent interacting with either Mouse 1 or Mouse 2 was measured as mentioned above.
  • Interaction time was scored using the automated AnyMaze software by trained, independent observers who were blind to the treatment group. [0189] Reciprocal Social Interaction Test.
  • Reciprocal social interaction test was performed as previously described (Buffington et al., 2016; Sgritta et al., 2019). Mice were placed in a 25 x 25 x 25 cm Plexiglas neutral arena with a novel conspecific matched mouse according to genotype, age, sex, and/or treatment. Trained, independent observers recorded the time a pair of mice socially interacted, using the AnyMaze software. Interactions included close following, touching, nose-to- nose sniffing, nose-to-anus sniffing, grooming and/or crawling over/under each other. The human observers were blind to the treatment group.
  • 16S rRNA Gene Sequencing was performed by the Center for Metagenomic & Microbiome Research at Baylor College of Medicine as previously described (Buffington et al., 2016; Sgritta et al., 2019). Methods were adapted from protocols developed for the NIH-Human Microbiome Project (Human Microbiome Project, 2012; Methe et al., 2012). Bacterial genomic DNA was extracted using MO BIO PowerSoil DNA Isolation Kit (MO BIO Laboratories).
  • the 16S rDNA V4 region was amplified using PCR with primers 515F (5 - GTGCCAGCMGCCGCGGTAA-3') and 806R (5 '-GGACT ACH VGGGTWTCTA AT-3 ') (Caporaso et al., 2011) and sequenced on the MiSeq platform (Illumina) using the 2x250 bp paired- end protocol yielding paired-end reads that overlap almost completely.
  • the primers used for amplification contained adapters for MiSeq sequencing and single-end barcodes allowing pooling and direct sequencing of PCR products.
  • the 16S rRNA gene read pairs were processed using QIIME2 v2020.6 (Bolyen et al., 2019). Reads were demultiplexed based on the unique molecular barcodes, trimmed to 220 base pairs in length for the forward reads and 140 base pairs for the reverse, denoised, merged, and filtered for chimeric reads with the DADA2 plugin (Callahan et al., 2016). 96.6% of the resulting amplicons were 252 or 253 base pairs in length, and amplicons less than 250 base pairs in length were discarded.
  • Taxonomic assignments were generated for the resulting amplicon sequence variants (AS Vs) by the taxonomic classifier in the DADA2 R package (version 1.10.0) using the SILVA database [version 138, (Quast et al., 2013)], with the option to add species-level information via exact matching where possible.
  • sequence variants were placed into the Greengenes 16S rRNA phylogeny using the SEPP QIIME2 plugin (Janssen et al.,
  • the median number of reads per sample before processing was 48,428 (minimum 9,445), and after processing, the median number of merged reads per sample was 32,224 (minimum 6,287).
  • genomic hidden states are predicted using castor (Louca and Doebeli, 2018), and MetaCyc pathway abundances (Caspi et al., 2020) are inferred using MinPath (Ye and Doak, 2009).
  • Lactobacillus reuteri 6475 was cultured anaerobically in MRS broth at 37°C in a 90% N2, 5% CO2, 5% Fh environment as previously described (Buffington et al., 2016). Briefly, cultures were centrifuged, washed, resuspended in PBS, and frozen at -80°C until use. PBS (vehicle) or L. reuteri was added to the drinking water daily to minimize dosage variability. The experimental group received live bacteria ( ⁇ 1 x 10 8 organisms/mouse/day), while the control group received equal volume of PBS. Mice drank the treated water ad libitum during the treatment period. Fecal sample collection, behavioral assays, tissue collection and electrophysiological recordings were initiated 4 weeks after the beginning of the treatment period.
  • Electrophysiological Recordings were performed as previously described (Buffington et al., 2016; Sgritta et al., 2019). Briefly, animals were anaesthetized with isoflurane and then decapitated. The brain was rapidly removed from the skull and fixed on a vibroslicer stage (VT lOOOS, Leica Microsystems, Buffalo Grove, IL) with cyanoacrylic glue.
  • VT lOOOS Leica Microsystems, Buffalo Grove, IL
  • Acute 220-250 pm-thick horizontal slices were cut in ice-cold (2-3 °C) cutting solution containing the following (in mM): 87 NaCl, 25 NaHCOs, 25 glucose, 75 sucrose, 2.5 KC1, 1.25 NaH 2 P0 4 , 0.5 CaCl 2 and 7 MgCh (equilibrated with 95% 0 2 -5% CO2 gas mixture, pH 7.3-7.5).
  • the slices were perfused with oxygenated ACSF (2 ml/min) containing the GABAA receptor antagonist picrotoxin (100 mM; Sigma- Aldrich, USA) and maintained at 32°C with a Peltier feedback device (TC-324B, Warner Instrument). [0196] Recordings were performed with Multiclamp700B (Molecular Devices), sampled at 20 kHz with Digidata 511440A (Molecular Devices) interface, filtered online at 3 kHz with a Bessel low-pass filter and analyzed off-line with pClamplO software (Molecular Devices).
  • Patch pipettes were pulled from borosilicate glass capillaries (World Precision Instruments, Inc., FL) and filled with following intracellular solution (in mM): 117 CsMeS0 3 , 0.4 EGTA, 20 HEPES, 2.8 NaCl, 2.5 Mg-ATP, and 0.25 Na-GTP; 5 TEA Cl, pH was adjusted to 7.3 and osmolarity to 290 mOsm using a vapor pressure osmometer Vapro5600 (ELITechGroupWescor, South Logan, Utah, USA). When filled with the intracellular solution, patch pipettes had a resistance of 2.0-3.0 MW before seal formation.
  • ventral tegmental area was visually identified by infrared differential interference contrast video microscopy and lateral VTA was identified considering the medial lemniscus and the medial terminal nucleus of the accessory optic tract as anatomical landmarks.
  • Dopaminergic (DA) neurons in this area were identified evaluating the following features: 1) cells firing at a frequency of 1-5 Hz and the spike width > 1 ms in cell attached configuration, 2) membrane capacitance (Cm) > 28 pF and 3) the presence of an Ih current and a leak current > 150 pA, when hyperpolarized from -40 mV to -120 mVin 10 mV steps (Bariselli etal., 2016; Huang etal., 2016; Zhang et al., 2010) . Passive electrode-cell parameters were monitored throughout the experiments, analyzing passive current relaxations induced by 10 Mv hyperpolarizing steps applied at the beginning of every trace. Variation of series resistance (Rs) > 20% led to the rejection of the experiment. AMPAR/NMDAR ratios were calculated as previously described (Buffington et al., 2016; Huang et al., 2016; Sgritta et al., 2019).
  • L2/3 neurons of the mPFC were identified under visual guidance and were clamped at -70 mV.
  • mEPSCs were recorded with a background activity protocol, in the presence of 100 pM picrotoxin and of 1 pM TTX. Recordings were low-pass filtered at 2 Hz and sampled at 20 kHz and analyzed off-line with pClamplO software. All spontaneous currents were digitally filtered at 1.5 kHz and analyzed off-line. Automated mEPSC analysis was performed with Clampfit software. A further visual inspection of detected signals allowed the inventors to reject noise artifacts. A period of 5 min was used to evaluate mEPSC frequency and mEPSC amplitude.
  • Fluorescent imaging and data acquisition were performed on a Zeiss Axio Imager.
  • Z2 microscope (Carl Zeiss Microimaging) mounted with an AxioCam digital camera (Carl Zeiss Microimaging). Images were captured using AxioVision acquisition software (Carl Zeiss Microimaging). All images within a given dataset were acquired at identical exposure times, within a given channel, to allow comparison of signal intensity.
  • Oxytocin Administration Oxytocin (Tocris Bioscience) was administrated intranasally at a dose of 200 pg/kg as previously reported (Buffington et al., 2016; Sgritta et al., 2019). Briefly, oxytocin was dissolved in 10% dimethyl sulfoxide (DMSO)/90% PBS. 10% DMSO/90% PBS was used as the vehicle control. 1.25 pL of vehicle or oxytocin were administrated into each nostril from a P10 pipette 30 minutes prior to social behavior testing, since the effects of oxytocin last 2 hours with this delivery method (Penagarikano et al., 2015).
  • DMSO dimethyl sulfoxide
  • Metabolomic Profiling Fecal samples were collected from mice in sterile microcentrifuge tubes, frozen immediately on dry ice, and stored at -80°C until processing. Samples were prepared using the automated MicroLab STAR system (Hamilton Co.) and analyzed using ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) on a Waters (Milford, MA) ACQUITY ultraperformance liquid chromatography (UPLC) and a Thermo Scientific Q-Exactive high resolution/accurate mass spectrometer interfaced with a heated electrospray ionization (HESI-II) source and Orbitrap mass analyzer operated at 35,000 mass resolution by Metabolon Inc. (Durham, North Carolina).
  • UPLC-MS/MS ultrahigh performance liquid chromatography-tandem mass spectrometry
  • HESI-II heated electrospray ionization
  • Orbitrap mass analyzer operated at 35,000 mass resolution by Metabolon Inc. (Durham,
  • Tetrahydrobiopterin (BH4) Administration (6R)-5,6,7,8-Tetrahydrobiopterin dihydrochloride (BH4, Sigma-Aldrich #T4425) was administered via oral gavage at a dose of 20 mg/kg. Briefly, BH4 was dissolved (5 mg/mL) in anaerobic water under anaerobic conditions (90% N2, 5% CO2, 5% 3 ⁇ 4 environment) to prevent oxidation. Aliquots were frozen at -80°C until use. Mice were gavaged with either vehicle or BH4 daily for 2-4 weeks and 2 hours prior to behavior tests.
  • L-371,257 (Tocris Bioscience) was administrated intranasally at a dose of -300 pg/kg, as previously reported (Penagarikano el al, 2015). Briefly, L-371,257 was dissolved in 10% dimethyl sulfoxide (DMSO) in PBS and 10% DMSO in PBS was used as the vehicle control. Two microliters of L-371,257 (or vehicle) were administrated into each nostril 20 minutes prior to reciprocal social interaction test.
  • DMSO dimethyl sulfoxide
  • SPRi3 (Tocris Bioscience) was administered intraperitoneally (I.P.) at a dose of 300 mg/kg based on previous reports (Cronin et al., 2018; Latremoliere el al., 2015).
  • Beta diversity PERMANOVA hypothesis testing was performed for all dissimilarity metrics using the adonis function in the R package vegan v2.5-6 (Oksanen et al., 2019), adjusting for sex, cage, and/or time covariates where applicable.
  • Alphadiversity indices were calculated using phyloseq, after subsampling sequence variant tables to the minimum number of reads assigned to a sample in a given experiment.
  • Hypothesis testing for alpha-diversity metrics was performed using the breakaway package v4.7.2 for richness estimates (Willis et al., 2020), and using linear mixed models for the Shannon index.
  • Neurohypophysial function and pteridines effect of (6R)-5,6,7, 8-tetrahydro-alpha-biopterin on bioassayed hypothalamo-neurohypophysial vasopressin and oxytocin in the rat.
  • OXTR oxytocin receptor gene
  • Oxytocin injected into the ventral tegmental area induces penile erection and increases extracellular dopamine in the nucleus accumbens and paraventricular nucleus of the hypothalamus of male rats.
  • the European journal of neuroscience 26, 1026-1035 The European journal of neuroscience 26, 1026-1035.
  • Caspr2 a new member of the neurexin superfamily, is localized at the juxtaparanodes of myelinated axons and associates with K+ channels. Neuron 24, 1037- 1047.
  • Nod2 is essential for temporal development of intestinal microbial communities. Gut 60, 1354-1362.

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Abstract

Selon des modes de réalisation, la divulgation concerne des méthodes et des compositions se rapportant à la manipulation du microbiome chez un sujet atteint au moins d'un trouble du comportement social, d'un dysfonctionnement du système congénital et/ou d'une anomalie de niveau de métabolite de bioptérine. Selon des modes de réalisation particuliers, le microbiome du sujet est manipulé en vue d'augmenter la bioptérine ou un métabolite de celle-ci. Selon des modes de réalisation précis, il est administré au sujet atteint au moins d'un trouble du comportement social, d'un dysfonctionnement du système congénital et/ou d'une anomalie de niveau de métabolite de bioptérine, une quantité efficace de bioptérine ou d'un métabolite de celle-ci pour la réduction d'au moins un symptôme du trouble du comportement social, du dysfonctionnement du système congénital et/ou de l'anomalie de niveau de métabolite de bioptérine. <i />
PCT/US2022/071003 2021-03-09 2022-03-07 Méthodes d'induction de bioptérine et de métabolites apparentés WO2022192854A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115273987A (zh) * 2022-09-23 2022-11-01 中国农业科学院农业质量标准与检测技术研究所 一种特征品质成分的识别方法、系统、设备及存储介质

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020116733A1 (fr) * 2018-12-07 2020-06-11 (주) 에이투젠 Nouvelle souche atg-f4 de lactobacillus reuteri possédant une fonction d'amélioration de la sécrétion de dopamine et composition pharmaceutique la comprenant pour la prévention ou le traitement de la psychopathie
WO2020160183A1 (fr) * 2019-01-29 2020-08-06 Holobiome, Inc. Procédés et compositions pour traiter et prévenir des troubles du système nerveux central et d'autres états provoqués par une dysbiose microbienne intestinale

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020116733A1 (fr) * 2018-12-07 2020-06-11 (주) 에이투젠 Nouvelle souche atg-f4 de lactobacillus reuteri possédant une fonction d'amélioration de la sécrétion de dopamine et composition pharmaceutique la comprenant pour la prévention ou le traitement de la psychopathie
WO2020160183A1 (fr) * 2019-01-29 2020-08-06 Holobiome, Inc. Procédés et compositions pour traiter et prévenir des troubles du système nerveux central et d'autres états provoqués par une dysbiose microbienne intestinale

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115273987A (zh) * 2022-09-23 2022-11-01 中国农业科学院农业质量标准与检测技术研究所 一种特征品质成分的识别方法、系统、设备及存储介质

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