WO2018013811A1 - Diagnostic et méthodes de traitement du syndrome de fatigue chronique et des troubles du spectre autistique - Google Patents

Diagnostic et méthodes de traitement du syndrome de fatigue chronique et des troubles du spectre autistique Download PDF

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WO2018013811A1
WO2018013811A1 PCT/US2017/041932 US2017041932W WO2018013811A1 WO 2018013811 A1 WO2018013811 A1 WO 2018013811A1 US 2017041932 W US2017041932 W US 2017041932W WO 2018013811 A1 WO2018013811 A1 WO 2018013811A1
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metabolites
subject
ceramide
suramin
metabolic pathway
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PCT/US2017/041932
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Robert K. Naviaux
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The Regents Of The University Of California
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/145—Amines having sulfur, e.g. thiurams (>N—C(S)—S—C(S)—N< and >N—C(S)—S—S—C(S)—N<), Sulfinylamines (—N=SO), Sulfonylamines (—N=SO2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A61K31/7072—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid having two oxo groups directly attached to the pyrimidine ring, e.g. uridine, uridylic acid, thymidine, zidovudine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2570/00—Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/30—Psychoses; Psychiatry
    • G01N2800/306—Chronic fatigue syndrome
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/50—Determining the risk of developing a disease

Definitions

  • the disclosure relates to biomarkers useful for diagnosing and predicting the development of chronic fatigue syndrome (CFS) .
  • CFS chronic fatigue syndrome
  • the disclosure further provides methods to reset metabolism and facilitate healing in CFS patients by administering antipurinergic compounds as well as the treatment of mitochondrial diseases and disorders with antipurinergic compounds.
  • Autism spectrum disorders affect 1-2% of children in the US.
  • Autism spectrum disorder is the name for a group of developmental disorders.
  • ASD includes a wide range, "a spectrum,” of symptoms, skills, and levels of disability. People with ASD often have these characteristics: ongoing social problems that include difficulty communicating and interacting with others; repetitive behaviors as well as limited interests or activities; symptoms that typically are recognized in the first two years of life; and symptoms that hurt the individual's ability to
  • CFS chronic fatigue syndrome
  • Chronic fatigue syndrome is a multi-system disease that can cause long-term pain and disability. It is difficult to diagnose because of its protean symptoms and the lack of a diagnostic laboratory test.
  • the disclosure demonstrates that targeted, broad- spectrum metabolomics of plasma not only revealed a characteristic chemical signature, but also revealed an unexpected underlying biology. Metabolomics showed that chronic fatigue syndrome is a highly concerted hypometabolic response to environmental stress that traces to mitochondria and was similar to the classically studied developmental state of dauer state of the nematode.
  • the disclosure provides methods to reset metabolism and facilitate healing in subject with mitochondrial disorders including autism spectrum disorders and chronic fatigue syndrome by administering anti-purinergic compounds such as suramin.
  • anti-purinergic compounds such as suramin.
  • antipurinergic compounds disclosed herein turn off the cell danger response (CDR) which is controlled by purinergic signaling such as the biological mechanism of chronic fatigue syndrome.
  • the disclosure provides methods of diagnosis.
  • the disclosure provides a method to determine a subject's risk of having or developing chronic fatigue syndrome (CFS) comprising: detecting an amount of each of a plurality of metabolites in a biological sample obtained from the subject, the plurality of metabolites comprising at least seven metabolites, each of the at least seven metabolites being in a metabolic pathway selected from the group of pathways consisting of: a sphingolipid metabolic pathway, a phospholipid metabolic pathway, a glycosphingolipid metabolic pathway, a purine metabolic pathway, a microbiome metabolic pathway, a cholesterol metabolic pathway, a vitamin B2 metabolic pathway, a pyrroline-5-carboxylic acid metabolic pathway, an arginine metabolic pathway, a proline metabolic pathway, and a branch chain amino acid pathway;
  • CFS chronic fatigue syndrome
  • determining the presence or absence of an alteration in the metabolic pathways of the subject based upon comparing the amounts of the detected metabolites of the subject versus the amounts of the metabolites detected from a control population that does not have CFS; and indicating that the subject has or is at risk of developing CFS based upon the determining that the metabolic pathways in the subject are altered in comparison to the control population.
  • the subject is a male subject
  • the method further comprises: detecting an amount of each of a plurality of metabolites in a biological sample obtained from the male subject, the plurality of metabolites comprising at least three metabolites, each of the at least 3 metabolites being in a metabolic pathway selected from the group of pathways consisting of: a serine/l-carbon metabolic pathway, a S-adenosyl methionine pathway, a S-adenosylhomocysteine metabolic pathway, a methionine metabolic pathway, a very long chain fatty acid oxidation metabolic pathway, a propiogenic amino acid metabolic pathway, and a threonine metabolic pathway; determining the presence or absence of an alteration in the metabolic pathways of the male subject based upon comparing the amounts of the detected metabolites from the male subject versus the amounts of the metabolites detected from a control population of male subjects that do not have CFS; and indicating that the male subject has or is at risk of developing CFS based
  • the metabolites of the male subject that are detected are phosphatidyl choline PC ( 16 : 0/16 : 0) , glucosylceramide
  • GC 18 : 1/16 : 0
  • P5C l-pyrroline-5-carboxylate
  • FAD flavin adenine dinucleotide
  • pyroglutamic acid 2-hydroxyisoccaproic acid
  • L-serine and lathosterol.
  • the subject is a female subject
  • the method further comprises: detecting an amount of each of a plurality of metabolites in a biological sample obtained from the female subject, the plurality of metabolites comprising at least three metabolites, each of the at least 3 metabolites being in a metabolic pathway selected from the group of pathways consisting of: a fatty acid oxidation metabolic pathway, a vitamin C/collagen metabolic pathway, a bile acid metabolic pathway; an endocannabinoid metabolic pathway, a vitamin B12 metabolic pathway; and an amino sugar metabolic pathway;
  • the metabolites that are detected are trihexosylceramide THC (18 : 1/24 : 0) , phosphatidyl choline
  • the metabolites that are detected are selected from group consisting of PC ( 16 : 0/16 : 0 ) , ceramide (dl8 : 1/24 : 2) , GC ( 18 : 1/16 : 0 ) , ceramide (dl 8 : 1/16 : 0 ) , THC (dl8 : 1/24 : 0) , PI(38:4), DHC (18:1/16:0) , PA (16:0/16:0) , 1- pyrroline-5-carboxylic acid, SM (dl8 : 1/24 : 2) ,
  • ceramide (dl8:l/16:l), ceramide (dl8: 1/18:0), ceramide (dl8: 1/26:2), ceramide (dl 8 : 1 /22 : 2 ) , L-serine, methionine sulfoxide,
  • the metabolites that detected are selected from the group consisting of ceramide (dl8 : 1/25 : 0) , THC (dl 8 : 1 /24 : 0 ) , PC ( 16 : 0/16 : 0 ) , lathosterol, hydroxyproline , PI ( 16 : 0/16 : 0 ) , ceramide (dl8 : 1/22 : 2) , adenosine, ceramide (dl8 : 1/24 : 2) , THC (dl8 : 1/16 : 0 ) , 2-octenoylcarnitine,
  • the metabolites from the subject are converted to a non-naturally occurring by-product that is analyzed.
  • the non-naturally occurring byproduct is a mass fragment.
  • the metabolites are detected by using one or more of the following: HPLC, TLC, electrochemical analysis, mass spectroscopy, refractive index spectroscopy (RI), Ultra-Violet spectroscopy (UV) ,
  • the biological sample is selected from the group consisting of cells, cellular organelles, interstitial fluid, blood, blood-derived samples, cerebral spinal fluid, and saliva.
  • the biological sample is a fluid sample.
  • the fluid sample is a serum sample.
  • the fluid sample is a urine sample.
  • the metabolites are detected by using at least mass spectroscopy.
  • metabolites are detected by using a combination of high performance liquid chromatography (HPLC) and mass spectroscopy (MS) .
  • HPLC high performance liquid chromatography
  • MS mass spectroscopy
  • each of the metabolites is measured based on a single run or injection.
  • the metabolites are detected by extracting from the biological sample each of the metabolites from each of the metabolic pathways.
  • an elevation or reduction in the detected amount of the metabolite from the subject of at least 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% in comparison to the amount of the same metabolite from the control population indicates an alteration in a metabolic pathway.
  • the disclosure further provides a method of treatment comprising: administering an antipurinergic compound to a subject that has been indicated as having CFS .
  • the antipurinergic compound is a P2Y inhibitor compound.
  • the P2Y inhibitor is suramin.
  • the subject has been indicated as having CFS based upon carrying out a metabolomics method disclosed herein .
  • the disclosure also provides a method for detecting the effectiveness of a treatment for a subject that has CFS comprising: carrying out a metabolomics method disclosed herein prior to treatment; carrying out the same metabolomics post treatment; and comparing the number of
  • the disclosure also provides a method of treating a subject suffering from an inherited or acquired mitochondrial disease or disorder comprising administering a therapeutically effective amount of an antipurinergic compound, e.g., suramin or derivative thereof. It is contemplated that administration of the an antipurinergic compound, e.g., suramin or derivative thereof. It is contemplated that administration of the an antipurinergic compound, e.g., suramin or derivative thereof. It is contemplated that administration of the
  • antipurinergic compound e.g., suramin or derivative thereof increases sulfur levels or free thiols in mitochondrial
  • the disclosure provides a method of treating a subject suffering from an inherited or acquired mitochondrial disease or disorder comprising administering a antipurinergic compound, e.g., suramin or derivative thereof.
  • a antipurinergic compound e.g., suramin or derivative thereof.
  • the mitochondrial disease or disorder is selected from the group consisting of bipolar disorder, multiple sclerosis, Parkinson's disease, schizophrenia, depression, autism, chronic fatigue syndrome, Friedreich's Ataxia, Leber's hereditary optic neuropathy, myoclonic epilepsy and ragged-red fibers (MERRF) , Mitochondrial encephalomyopathy, mitochondrial myopathy, mitochondrial neurogastrointestinal encephalopathy
  • MNGIE lactic acidosis
  • MELAS stroke-like syndrome
  • Kearns- Sayre syndrome Pearson marrow syndrome
  • NARP neuropathy ataxia and retinitis pigmentosa
  • POLG Polymerase gamma disorders including but not limited to Alpers-Huttenlocher syndrome and ataxia neuropathy spectrum (ANS) disorders
  • ANS neuropathy spectrum
  • other mitochondrial DNA depletion syndromes including but not limited to those caused by gene defects in TK2, Twinkle, DGUOK, RRMB2, SUCLA2, SUCLG1, MPV17, ANT1, MFN1, MFN2, and OPA1, subacute necrotizing
  • mitochondrial diseases include neurogenic muscle weakness, ataxia and retinitis pigmentosa (NARP), progressive external opthalmoplegia (PEO) , and Complex I disease, Complex II disease, Complex III disease, Complex IV disease and Complex V disease, which relates to dysfunction of the OXPHOS complexes, and MEGDEL syndrome ( 3-methylglutaconic aciduria type IV with sensorineural deafness, encephalopathy and Leigh-like
  • antipurinergic compound e.g., suramin or derivative thereof is about lOOmg to 1 g per day (e.g., about 10-20 mg/kg) ; typically administered weekly.
  • the suramin can be formulated as an immediate release or a delayed or extended release form.
  • the suramin can be formulated as a delayed or controlled release dosage form that provides increased delivery to the small intestine.
  • the formulation can comprise an enteric coating that releases the suramin or derivative thereof when the suramin or derivative thereof reaches the small intestine or a region of the
  • the coating can be selected from the group consisting of polymerized gelatin, shellac, methacrylic acid copolymer type CNF, cellulose butyrate phthalate, cellulose hydrogen phthalate, cellulose proprionate phthalate, polyvinyl acetate phthalate (PVAP) , cellulose acetate phthalate (CAP) , cellulose acetate trimellitate (CAT) , hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate, dioxypropyl methylcellulose succinate, carboxymethyl ethylcellulose (CMEC) , hydroxypropyl methylcellulose acetate succinate (HPMCAS) , and acrylic acid polymers and copolymers, typically formed from methyl acrylate, ethyl acrylate, methyl methacrylate and/or ethyl
  • PVAP polyvinyl acetate phthalate
  • CAP cellulose acetate phthalate
  • CAT cellulose acetate trimellitate
  • composition can be administered orally or parenterally.
  • the subject is under oxidative stress. In another embodiment, the subject has decreased thiol levels compared to a non-affected subject.
  • the administering results in improvement in mitochondrial activity markers compared to levels before administration of the suramin or derivative thereof.
  • Exemplary mitochondrial activity markers include, but are not limited to, free thiol levels, glutathione (GSH) , reduced
  • GSSH glutathione
  • AOPP advanced oxidation protein products
  • FRAP ferric reducing antioxidant power
  • GSSH glutathione
  • AOPP advanced oxidation protein products
  • FRAP ferric reducing antioxidant power
  • lactic acid, pyruvic acid, lactate/pyruvate ratios phosphocreatine, NADH (NADH+H + ) or NADPH (NADPH+H + ) , NAD or NADP levels
  • ATP anaerobic threshold
  • reduced coenzyme Q oxidized coenzyme Q
  • total coenzyme Q oxidized cytochrome C, reduced cytochrome C, oxidized cytochrome C/reduced cytochrome C ratio, etc.
  • the administering results in increased thiol levels compared to levels before administration of the suramin or derivative thereof.
  • the suramin or derivative thereof further comprises a pharmaceutically acceptable carrier. It is further contemplated that the suramin or derivative product is formulated as a sterile pharmaceutical composition.
  • a suramin or derivative product or composition is administered with a second agent useful to treat inherited or acquired mitochondrial diseases or disorders.
  • second agents include, but are not limited to, cysteamine, cystamine, coenzyme Q10, coenzyme Q10 analogs, idebenone, decylubiquinone , Epi-743, resveratrol and analogs thereof, arginine, vitamin E, tocopherol, MitoQ,
  • the subject is a child or
  • the methods of the disclosure also include use of a suramin or derivative product in preparation of a
  • medicament for treatment an inherited or acquired mitochondrial disease and use of a suramin or derivative product in preparation of a medicament for administration in combination with a second agent for treating an inherited or acquired mitochondrial disease. Also included is use of a second agent for treating an inherited or acquired mitochondrial disease.
  • Figure 1A-E presents a metabolomic diagnosis of chronic fatigue syndrome.
  • A Males
  • B Females.
  • Multivariate analysis using partial least squares discriminant analysis (PLSDA) clearly distinguished controls and patients with chronic fatigue in both males and females.
  • PLSDA partial least squares discriminant analysis
  • C Males, the top five pathway disturbances in males were responsible for 82% of the metabolic impact. These were sphingolipids (49%),
  • Figure 2A-B provides for the characterization of total, low, and high metabolite abnormalities in chronic fatigue syndrome.
  • Metabolites with Z-scores ⁇ 2.0 are indicated in red. Metabolites with Z-scores ⁇ -2.0 are indicated in green. Both the total number of metabolite abnormalities and the number of metabolites that were decreased were significantly increased in patients with chronic fatigue syndrome.
  • Figure 3A-B provides a rank order of distinguishing metabolite abnormalities in chronic fatigue syndrome.
  • Figure 4A-B presents a cytoscape visualization of
  • Figure 5A-B provides the distribution of diagnostic and personalized metabolic abnormalities in chronic fatigue syndrome.
  • Figure 6A-B presents the diagnostic performance of targeted metabolomics in chronic fatigue syndrome.
  • AUROC Receiver Operator Characteristic
  • metabolites were selected as a diagnostic classifier in females as described above.
  • the 13 metabolites were trihexosylceramide
  • THC (18 : 1/24 : 0) , phosphatidyl choline PC ( 16 : 0/16 : 0 ) , hydroxyproline , ceramide (dl 8 : 1 /22 : 2 ) , lathosterol, adenosine, phosphatidylinositol PI (16: 0/16: 0) , flavin adenine dinucleotide (FAD), 2- octenoylcarnitine , phosphatidyl choline plasmalogen PC (22 : 6/P18 : 0) , phosphatidyl choline PC (18 : 1/22 : 6) , l-pyrroline-5-carboxylate
  • Figure 7 displays the mitochondrial control of redox, NADPH, nucleotide, and methylation Pathways. In embryonic cells and cancer, MTHFD2L is expressed and one-carbon units are
  • Cytosolic Trifunctional Cl-THF Synthase [12—Formyl-THF Synthase, 13—Methenyl-THF Cyclohydrolase, 14--NADPH-dependent Methylene-THF Dehydrogenase], 15--Formyl-THF Dehydrogenase, 16-- Homocysteine Methyl Transferase (Methionine Synthase, CblG) . 17-- Methionine Adenosyl Transferase (MAT) . 18*---Multiple DNA- , RNA-, Protein--, Neurotransmitter, and Other Methyltrans ferase reactions in the nucleus, cytosol, and mitochondria. 19--S-Adenosyl
  • SAHH Homocysteine Hydrolase
  • CBS 20--Cystathionine ⁇ -Synthase
  • CBS 21—Cystathionase (Cystathionine ⁇ -lyase)
  • GCS GCS
  • GCS GCS
  • Diphosphate Kinase 25--ATP Synthase (Complex V), 26--Propionyl CoA Carboxylase, 27--Methylmalonyl CoA Mutase, 28--Betaine Homocysteine Methyltrans ferase , 29--Choline Dehydrogenase, 30--Betaine Aldehyde Dehydrogenase, 31--S-Adenosylmethionine decarboxylase (adoMetDC, AMD1, SAMDC) , 32--Spermidine synthase, 33--Spermine synthase, 34--- Methylthioadenosine phosphorylase (MTAP) , 35--Methionine synthase reductase (MSR, MTRR, CblE) , 36--Delta Amino Levulinic Acid
  • FIG. 8A-B Principal Components Analysis
  • Figure 9 provides a CONSORT flow chart of a suramin study design overview, including study recruitment, allocation and analysis.
  • the two treatment groups were well matched by the 3 parameters that were the basis of the pairing (age, ADOS, and IQ) , and were incidentally well matched for 7 other anthropometric criteria (see Table 14) .
  • Twenty families were interviewed. Four children were excluded for cause; two were taking prescription medications, one was 18 years old, and one lived 2.5 hours away. Sixteen children were eligible for enrollment. Ten children were matched into 5 pairs on the basis of age, non-verbal Leiter-3 IQ, and ADOS-2 scores. One child in each pair was then randomized to receive either saline or suramin (see also Table 14) .
  • Figure 10 provides a diagram of the suramin study design. Phone interviews, parent journals, and clinical
  • Figure 11A-I presents the results of safety monitoring with suramin administration.
  • Suramin Safety Monitoring A: No change was observed in Cortisol levels between saline and suramin treated infusion and decrease in the afternoon post-infusion were seen in both saline and suramin treated subjects.
  • B Proteinuria was unchanged by suramin treatment
  • C Creatinine was unchanged by suramin by 2-way ANOVA, but was normalized after 6-weeks in a paired analysis (see Figure 8B) .
  • D Hemoglobin subjects.
  • Figure 12A-D presents suramin pharmacokinetics.
  • A 2- compartment model of suramin blood concentrations. The first 48 hours were dominated by the distribution phase. Over 90% of the model is described by the elimination phase.
  • B Plasma suramin concentrations
  • C A 2-compartment model correlated well with measured values
  • D Pediatric PK parameters of suramin.
  • Figure 13A-B presents suramin pharmacometabolomics .
  • Figure 14 presents suramin pharmacometabolomics.
  • Figure 15A-B provides for the visualization of the suramin pharmacometabolomics pathway.
  • A After 2 days.
  • B After 6 weeks. Metabolites indicated in red are increased, and those in green are decreased compared to controls (see z-score scale in upper right) .
  • Figure 16A-dd demonstrates the various outcomes with suramin treatment versus placebo.
  • A 6-week ADOS Comparison Scores by 2-Way ANOVA.
  • B 6-Week ADOS Comparison Score Improvement after suramin.
  • C 6-Week ADOS Social Affect Score Improvement after suramin.
  • D 6-Week ADOS Restricted and Repetitive Behavior Score Improvement after Suramin.
  • E 2-Day ADOS Comparison Scores were not changed.
  • F No change in 6-Week ADOS Scores in subjects receiving saline placebo.
  • G No change in 6-Week ADOS Social Affect Scores in subjects receiving placebo.
  • H No change in 6- Week ADOS Restricted and Repetitive Behavior Scores in subjects receiving placebo.
  • ATEC speech, language, and communication scores 6-weeks after suramin Improved ATEC speech, language, and communication scores 6-weeks after suramin.
  • U No change in 7- day ATEC speech, language, and communication after saline.
  • V No change in 7-day ATEC sociability after saline.
  • W No change in 6- week ATEC speech, language, and communication scores 6-weeks after saline
  • Y No change in 2-day ADOS scores after suramin.
  • Figure 19 shows that hyperactivity in male ASD mice was normalized by antipurinergic therapy with suramin. Two-way ANOVA with Bonferroni post hoc testing of center region entries in the first 10 minutes of exploration in the open field test. Suramin had no effect on the activity of normal control animals.
  • Figure 20 demonstrates that antipurinergic therapy restores autonomic balance and normalizes stress-induced
  • SIH hyperthermia
  • Figure 21A-B shows the increased SIH Response in ASD mice is the result of a decreased basal body temperature and that treatment with antipurinergic therapy with suramin is corrective.
  • A The post stress temperatures (dark bars) taken at 10 minutes after the baseline temperatures were not different. However, the pre-stress basal body temperature of the Poly ( IC) -exposed controls (PIC-SAL; white bars) was significantly lower than the basal temperatures of the other 3 experimental groups.
  • PIC-SAL pre-stress basal body temperature of the Poly
  • FIG. 22A-B demonstrates that a maternal immune activation model of autism produces a state of chronic mild hypothermia.
  • the basal temperature of animals 1 hour after a purinergic stimulus given by an injection of ATP (0.5 mL of 25 mM ip) is 1.8 °C lower than saline injected controls.
  • Figure 23 demonstrates that a single-dose of suramin corrected a short-term memory deficit in the Poly(IC) Mouse Model of ASD. A single dose of suramin given to 5 month-old animals corrected the memory deficit in this model.
  • Figure 24 demonstrates that a single-dose of suramin corrected social abnormalities in a Poly(IC) mouse model of ASD. A single dose of suramin given to 5 month-old animals corrected the social approach abnormalities in this model.
  • small molecules includes organic and inorganic molecules, such as those present in a biological sample obtained from a patient or subject.
  • examples of small molecules include sugars, fatty acids, amino acids, nucleotides, intermediates formed during cellular processes, and other small molecules found within a cell.
  • the small molecules are metabolites.
  • a small molecule is a chemical compound, e.g., a drug.
  • CNVs chromosomal copy number variants
  • CFS Chronic fatigue syndrome
  • Diagnosis requires that the patient have the following three symptoms :
  • CFS chronic fatigue syndrome
  • disorder as used herein is intended to be generally synonymous, and is used interchangeably with, the terms “disease,” “syndrome,” and “condition” (as in medical condition), in that all reflect an abnormal condition of the human or animal body or of one of its parts that impairs normal functioning, is typically manifested by distinguishing signs and symptoms.
  • the disclosure provides, for example, a method of treating and/or preventing autism, autism-like disorders, and/or symptoms using suramin and derivatives thereof as therapeutic agents.
  • Autism and autism-like disorders include, for example, familial forms of autism, mental retardation, and/or Asperger syndrome.
  • Other disorders that can be treated with the compositions and methods described herein are mitochondrial disorders and chronic fatigue syndrome .
  • Mitochondrial disorders or "-dysfunction” are defects in oxidative metabolism and are found in many chronic illnesses including bipolar disorder, multiple sclerosis, Parkinson's disease, schizophrenia, depression, autism, and chronic fatigue syndrome. Elevated levels of reactive oxygen and nitrogen species together with elevated pro-inflammatory cytokines and reduced levels of glutathione impair oxidative metabolism result in mitochondrial dysfunction.
  • Chronic fatigue syndrome is a complex and is characterized by profound fatigue and disability, episodes of cognitive dysfunction, sleep disturbance, autonomic
  • small molecule metabolite profile refers to the composition, amounts, and/or identity, of small molecule
  • the small molecule metabolite profile provides information related to the metabolism or metabolic pathways that are active in a cell, tissue or organism.
  • the small molecule metabolite profile provides data for developing a "metabolomic profile" (also referred to as “metabolic profile") of active or inactive metabolic pathways in a cell, tissue, or subject.
  • the small molecule metabolite profile includes, e.g., the quantity and/or type of small molecules present.
  • metabolite profile can be obtained using a single measurement technique (e.g., HPLC) or a combination of techniques (e.g., HPLC and mass spectrometry) .
  • HPLC high-density lipoprotein
  • mass spectrometry mass spectrometry
  • a "metabolomic profile” is a profile of pathway activity associated with the small molecule metabolites.
  • the activity of the pathways is an indication of metabolic health.
  • one or more small molecule metabolites can be measured in a specific pathway, the small molecule metabolites can include intermediates as well as the end product.
  • the metabolomics profile identifies the pathway's "activity". If the pathway produced a normal amount of the metabolite, then the pathway is normal, however, if the pathway produces excessive or reduced amounts then the pathway has aberrant activity.
  • a disease state (or risk thereof) is identified by a plurality of aberrant pathways in a metabolomics profile.
  • the pathway can be identified numerically, by color, by code or other symbols as being aberrant or normal.
  • a small molecule metabolite profile and metabolomic profile can be obtained for a normal control (e.g., a "control small molecule metabolite profile” or “control metabolomic profile”) and would include an inventory of small molecules or metabolomic pathways that are active in similar cells, tissue or sample from a population of subject that are considered "normal” or "healthy”
  • control small molecule metabolite profile or "control metabolomic profile” would include the inventory and amounts of small molecules present (or metabolic pathways active) in, e.g., 70%, 80%, or 90%, but typically greater than 95% of a population that does not have any symptoms of CFS .
  • subject refers to an animal, including, but not limited to, a primate (e.g., human, monkey, chimpanzee, gorilla, and the like), rodents (e.g., rats, mice, gerbils, hamsters, ferrets, and the like), lagomorphs, swine (e.g., pig, miniature pig), equine, canine, feline, and the like.
  • a mammalian subject can refer to a human patient.
  • Suramin is a polysulfonated naphthylurea .
  • Suramin derivatives and analogues for use in the disclosed methods are also provided. Derivatives of suramin are known in the art (See U.S. Pat. No. 5,173,509, Braddock, P S., et al . 1994; Dhar, S., et al. 2000; Firsching, A., et al . 1995; Gagliardi, A. R. T., et al . 1998; Kreimeyer, A., et al . 1998; Marchetti, D., et al . 2003;
  • Suramin has many actions. One of its best-studied actions is as an inhibitor of purinergic signaling. It is the oldest member of a growing class of
  • antipurinergic drugs in development. Suramin was first synthesized in 1916, making it one of the oldest manmade drugs still in medical use. It is used to treat African sleeping sickness (trypanosomiasis), and remains on the World Health
  • the term "therapeutically acceptable” refers to those compounds (or salts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitable for use in contact with the tissues of subjects or patients without excessive toxicity, irritation, allergic response, immunogenicity, are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use.
  • mitochondrial-associated diseases and disorders including, but not limited to, autism spectrum disorders and chronic fatigue syndrome.
  • the disclosure also provides metabolomics diagnostics useful to diagnosing mitochondrial diseases and disorders
  • autism spectrum disorders including, but not limited to, autism spectrum disorders and chronic fatigue syndrome.
  • the disclosure provides for novel treatment options for to prevent or retard neuronal loss and/or improve short-term memory in a subject comprising administering to a subject in need thereof, a composition comprising an effective amount of a purinergic antagonist such as suramin or a derivative thereof.
  • Purinergic antagonist are a new class of drugs that can be used safely for the treatment of autism and related spectrum disorders like chronic fatigue syndrome, fibromyalgia, obsessive compulsive disorder (OCD) , generalized anxiety disorder (GAD) , bipolar depression, schizophrenia, subacute therapy of Traumatic Brain Injury (TBI), post-traumatic stress disorder (PTSD) , Chronic Traumatic
  • CTE Cerephalopathy
  • ADHD attention deficit hyperactivity disorder
  • ADD attention deficit hyperactivity disorder
  • Alzheimer dementia early Alzheimer dementia for the prevention of neuronal loss and improvement of short-term memory.
  • suramin The only non-cardiovascular, antipurinergic drug currently approved for human use is suramin. More than a dozen others are in development around the world for a variety of indications, but none for mitochondrial diseases and disorders or for autism spectrum disorders or chronic fatigue syndrome. Suramin has been used to treat African sleeping sickness for nearly 100 years at a "medium dose” designed to produce blood levels of 50-100 ⁇ for 1-3 months. This is generally well-tolerated, but side- effects can occur, particularly in fragile or malnourished patients. Since the 1990s, suramin has been used in adjunct cancer chemotherapy protocols at a "high-dose" designed to produce blood levels of 150-270 ⁇ for 3-6 months.
  • High-dose regimens produce side effects in up to 10% of subjects per month, so that by 5 months, over half of patients has reported at least one side effect. These include adrenal insufficiency, anemia, and peripheral neuropathy. This disclosure teaches how "low-dose" suramin can be used safely to treat mitochondrial disorders, autism, chronic fatigue syndrome and related disorders.
  • pathophysiologically related disorders including but not limited to: Autism, Chronic fatigue syndrome, Fibromyalgia, Obsessive compulsive disorder, Generalized anxiety disorder, Schizophrenia, Bipolar Depression, Subacute therapy for Traumatic Brain Injury
  • TBI Post-Traumatic Stress Disorder
  • CTE Chronic Traumantic Encephalopathy
  • ADHD Disabling Attention Deficit Hyperactivity Disorder
  • ADHD Alzheimer Disease
  • a subject to be treated by a compound disclosed herein has a developmental disorder.
  • a subject to be treated by a compound disclosed herein has a mental disorder.
  • the subject is an infant, child or adolescent.
  • the compounds of the disclosure can be formulated for deliver by admixture with pharmaceutically acceptable non-toxic excipients or carriers.
  • pharmaceutically acceptable salts of the addition salts with inorganic or organic acids (such as acetate, trifluoroacetate, propionate, succinate, benzoate, fumarate, maleate, oxalate, methanesulphonate , isethionate, theophyllinacetate , salicylate, methylenebis-p-oxynaphthoate, hydrochloride, sulphate, nitrate and phosphate) , the salts with alkali metals (sodium, potassium or lithium) or with alkaline-earth metals (calcium or magnesium) , the ammonium salt or the salts of nitrogenous bases (ethanolamine, trimethylamine , methylamine, piperidine, benzylamine, iV-benzyl-a- phenethylamine ,
  • the disclosure provides pharmaceutical compositions of an antipurinergic agent or their salts.
  • the antipurinergic agent or their physiologically acceptable salts or solvates may be formulated for administration for injection, or for oral, topical, nasal, inhalation, insufflation (either through the mouth or the nose) buccal, parenteral, rectal administration or other forms of administration.
  • the disclosure provides pharmaceutical compositions comprising effective amounts of antipurinergic agent together with pharmaceutically acceptable diluents, preservatives, solubilizers , emulsifiers, adjuvants, excipients and/or carriers.
  • compositions include diluents of various buffer content (e.g., Tris-HCl, acetate, phosphate) , pH and ionic strength; additives such as detergents and solubilizing agents (e.g., Tween 80,
  • polysorbate 80 polysorbate 80
  • anti-oxidants e.g., ascorbic acid, sodium metabisulfite
  • preservatives e.g., thimerosal, benzyl alcohol
  • bulking substances e.g., lactose, mannitol
  • the antipurinergic agent is suramin or a derivative thereof .
  • compositions may also be incorporated into
  • Biocompatible absorbable polymers may be selected from the group consisting of aliphatic polyesters, copolymers and blends, which include, but are not limited to, homopolymers and copolymers of lactide (which include D-, L-, lactic acid and D-, L- and meso lactide) , glycolide (including glycolic acid) , epsilon- caprolactone , p-dioxanone (1, 4-dioxan-2-one) , alkyl substituted derivatives of p-dioxanone (i.e., 6, 6-dimethyl-l, 4-dioxan-2-one) , triethylene carbonate (1, 3-dioxan-2-one) , alkyl substituted derivatives of 1 , 3-dioxanone , delta-valerolact
  • compositions may influence physical state, stability, rate of in vivo release, and rate of in vivo clearance. See, e.g., Remington s Pharmaceutical Sciences, 18th ed. , (1990, Mack Publishing Co., Easton, Pa. 18042) pages 1435-1712).
  • the compositions may be prepared in liquid form, or be in dried powder, such as lyophilized form.
  • Solid dosage forms include tablets, capsules, pills, troches or lozenges, cachets or pellets.
  • liposomal or proteinoid encapsulation may be used to formulate compositions. Liposomal encapsulation may be used and the liposomes may be derivatized with various polymers. A description of possible solid dosage forms for the therapeutic is given by Marshall, K. In:
  • the formulation will include an antipurinergic agent (e.g., suramin and/or derivatives thereof) and inert ingredients (which allow for protection against the stomach environment and release of the biologically active material in the intestine) .
  • an antipurinergic agent e.g., suramin and/or derivatives thereof
  • inert ingredients which allow for protection against the stomach environment and release of the biologically active material in the intestine
  • a coating impermeable to at least pH 5.0 is useful.
  • examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT) , hydroxypropylmethylcellulose phthalate (HPMCP) , HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP) , Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP) , Eudragit L, Eudragit S, and Shellac. These coatings may be used as mixed films.
  • a coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings that make the tablet easier to swallow.
  • Capsules may consist of a hard shell (such as gelatin) for delivery of dry therapeutic, i.e., powder; for liquid forms, a soft gelatin shell may be used.
  • the shell material of cachets may be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet triturates, moist massing techniques can be used.
  • the therapeutic can be included in the formulation as fine multi-particulates in the form of granules or pellets.
  • the formulation of the material for capsule administration can also be as a powder, lightly compressed plugs or even as tablets.
  • the therapeutic can also be prepared by compression.
  • Colorants and flavoring agents may all be included.
  • the peptide (or derivative) may be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage
  • these diluents or fillers can include carbohydrates, especially mannitol, anhydrous lactose, cellulose (e.g., microcrystalline cellulose), sucrose, calcium hydrogen phosphate modified dextrans and starch.
  • Certain inorganic salts may also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride.
  • Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell .
  • Disintegrants may be included in the formulation of the therapeutic into a solid dosage form. Materials used as
  • disintegrates include, but are not limited to, starch (e.g., potato starch or the commercial disintegrant based on starch, Explotab) .
  • starch e.g., potato starch or the commercial disintegrant based on starch, Explotab
  • Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose , ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used.
  • Another form of the disintegrants are the insoluble cationic exchange resins.
  • Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants .
  • Binders may be used to hold the therapeutic agent together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch (e.g., pre-gelatinized maize starch) and gelatin. Others include methyl cellulose (MC) , ethyl cellulose (EC) and carboxymethyl cellulose (CMC) . Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) can both be used in alcoholic solutions to granulate the therapeutic.
  • natural products such as acacia, tragacanth, starch (e.g., pre-gelatinized maize starch) and gelatin. Others include methyl cellulose (MC) , ethyl cellulose (EC) and carboxymethyl cellulose (CMC) .
  • MC methyl cellulose
  • EC ethyl cellulose
  • CMC carboxymethyl cellulose
  • PVP polyvinyl pyrrolidone
  • HPMC hydroxy
  • An anti-frictional agent may be included in the formulation of the therapeutic to prevent sticking during the formulation process.
  • Lubricants may be used as a layer between the therapeutic and the die wall, and these can include but are not limited to; stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE) , liquid paraffin, vegetable oils and waxes, talc and silica. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000.
  • Glidants that can improve the flow properties of the drug during formulation and to aid rearrangement during compression can be added.
  • the glidants can include starch, talc, pyrogenic silica and hydrated silicoaluminate .
  • a surfactant can be added as a wetting agent.
  • Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium
  • Cationic detergents can be used and can include benzalkonium chloride or benzethomium chloride.
  • the list of potential non-ionic detergents that can be included in the formulation as surfactants are lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty cid ester, methyl cellulose and carboxymethyl cellulose. These surfactants can be present in the formulation of the protein or derivative either alone or as a mixture in different ratios.
  • Additives that potentially enhance uptake of the agent are, for example, the fatty acids oleic acid, linoleic acid and linolenic acid.
  • Controlled release oral formulation may be desirable.
  • the agent can be incorporated into an inert matrix that permits release by either diffusion or leaching mechanisms, e.g., gums. Slowly degenerating matrices may also be incorporated into the formulation. Some enteric coatings also have a delayed release effect .
  • coatings may be used for the formulation. These include a variety of sugars that can be applied in a coating pan.
  • the therapeutic agent can also be given in a film coated tablet and the materials used in this instance are divided into two groups. The first are the nonenteric materials and include methyl
  • the second group consists of the enteric materials that are commonly esters of phthalic acid.
  • a mix of materials can be used to provide the optimum film coating.
  • Film coating may be carried out in a pan-coater or in a fluidized bed or by compression coating.
  • Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use.
  • Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid) .
  • the preparations may also contain buffer salts, flavoring, coloring and sweetening agents as appropriate.
  • the compounds e.g., antipurinergic agents such as suramin and/or derivatives thereof
  • parenteral administration may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative.
  • the compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • the compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
  • the compounds disclosed herein may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection.
  • the compounds disclosed herein may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a soluble salt.
  • compositions may, if desired, be presented in a pack or dispenser device that may contain one or more unit dosage forms containing the active ingredient.
  • the pack may for example comprise metal or plastic foil, such as a blister pack.
  • the pack or dispenser device may be accompanied by instructions for
  • LD 50 the dose lethal to 50% of the population
  • ED 50 the dose therapeutically effective in 50% of the
  • the dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD 50 /ED 50 . While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects .
  • the data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans.
  • the dosage of such compounds lies within a range of circulating concentrations that include the ED 50 with little or no toxicity.
  • the dosage may vary within this range depending upon the dosage form employed and the route of administration utilized.
  • a dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC 50 (i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.
  • a compound disclosed herein and components of a therapeutic composition may be introduced parenterally, topically, or transmucosally, e.g., orally, nasally, or rectally, or
  • Parenteral administration includes, for example, intravenous injection, intra-arteriole , intramuscular, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration .
  • the compounds are suitable for oral, parenteral or intravenous administration.
  • the compound can be modified or otherwise altered so that it can effectively cross or be transported across the blood brain barrier.
  • Many strategies known in the art are available for molecules crossing the blood-brain barrier, including but not limited to, increasing the hydrophobic nature of a molecule; introducing the molecule as a conjugate to a carrier, such as transferring, targeted to a receptor in the blood-brain barrier, or to
  • a compound of the disclosure can be administered intracranially or intraventricularly.
  • osmotic disruption of the blood-brain barrier can be used to effect delivery of the compound to the brain (Nilayer et al., Proc. Natl. Acad. Sci. USA 92:9829-9833 (1995)).
  • a compound of the disclosure can be administered intracranially or intraventricularly.
  • osmotic disruption of the blood-brain barrier can be used to effect delivery of the compound to the brain (Nilayer et al., Proc. Natl. Acad. Sci. USA 92:9829-9833 (1995)).
  • a compound of the disclosure can be administered intracranially or intraventricularly.
  • osmotic disruption of the blood-brain barrier can be used to effect delivery of the compound to the brain (Nilayer et al., Proc. Natl. Acad. Sci. USA 92:9829-9833 (1995)).
  • a compound of the disclosure can be
  • liposomes administered in a liposome targeted to the blood-brain barrier.
  • Administration of pharmaceutical agents in liposomes are known (see Langer, Science 249:1527-1533 (1990); Treat et al., Liposomes in the Therapy of infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. pp. 317-327 and 353-365 (1989).
  • a therapeutic formulation comprising an antipurinergic agent can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss: New York, pp. 317- 327 and 353-365 (1989) ) .
  • a therapeutic formulation comprising a compound of the disclosure can be delivered in a controlled release system.
  • the antipurinergic agent may be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration.
  • a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al . , N. Engl. J. Med.
  • polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Press: Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds. ) , Wiley: New York (1984); Ranger and Peppas, J.
  • a controlled release system can be placed in proximity of the therapeutic target, e.g., the brain, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138
  • any of the materials described herein can be administered to any part of the mammal's body including, without limitation, brain, spinal fluid, blood stream, lungs, nasal cavity, intestines, stomach, muscle tissues, skin, peritoneal cavity, and the like.
  • a compound e.g., an antipurinergic agent
  • an aerosol preparation can be given to a mammal by inhalation.
  • duration of treatment with the materials described herein can be any length of time from as short as one day to as long as a lifetime (e.g., many years) .
  • a formulation comprising a compound of the disclosure can be administered once (or twice, three times, etc.) daily, weekly, monthly, or yearly.
  • Preparations for administration can include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.
  • non-aqueous solvents include, without limitation, propylene glycol, polyethylene glycol, vegetable oils, and injectable organic esters.
  • Aqueous carriers include, without limitation, water as well as alcohol, saline, and buffered solutions. Preservatives, flavorings, and other additives such as, for example, antimicrobials, anti-oxidants , chelating agents, inert gases, and the like may also be present.
  • a compound of the disclosure is administered in a dose sufficient to provide a therapeutically effective amount to an individual to provide a beneficial effect (e.g., to treat a symptom of mitochondrial disorders, autism spectrum disorders or chronic fatigue syndrome) of a subject.
  • a therapeutically effective dose of a compound of the disclosure can be determined empirically and depends on the type of treatment, the route of administration, and the size, weight, age and overall health of the patient, as is within the skill of one in the art such as a medical practitioner.
  • an antipurinergic compound e.g., suramin and/or derivatives thereof
  • the amount of an antipurinergic compound (e.g., suramin and/or derivatives thereof) disclosed herein which is administered as a unit dose will depend upon the type of pharmaceutical composition being administered, for example, a solution, a suspension, a gel, a film, an emulsion, a powder, or a sustained- release formulation.
  • the quantity of formulation needed to deliver the desired dose will also depend on the concentration of the compound of the disclosure in the composition. Such determinations are within the skill of one in the art.
  • an antipurinergic compound disclosed herein in the pharmaceutical compositions used in the methods of the disclosure will depend on a number of factors such as the chemical composition and/or modification of the compound, its bioavailability by the chosen route of administration, its efficacy, the desired frequency of administration combined with the desired single dosage of the formulation and whether the compound is administered in combination with other active agent (s) .
  • the dosage of a compound disclosed herein will be chosen to maximize cognitive functions of a subject.
  • Pharmacological data can be obtained from animal models and clinical trials with normal human volunteers or patients by one with skill in the art.
  • antipurinergic compound e.g., suramin and/or derivative thereof
  • dosages used for administration can include, but are not limited to, an effective amount within the dosage range of about 1-lOOmg per dose, typically about lOOmg to 1 g per day (e.g., about 10-20 mg/kg) .
  • Dosages can be administered in a single dose or in multiple doses, for example, dosages can be administered two, three, four, up to ten times daily depending on the type of treatment as well as on individual susceptibility. Dosages can be administered in a sustained release formulation which may allow for a compound disclosed herein to be administered less frequently such as six times a week, five times a week, four times a week, three times a week, twice a week, or once a week, once a month, once every two months, three months, four months, five months or six months or more. Infrequent administration can be accomplished by sustained release formulations.
  • compositions comprising an antipurinergic compound such as suramin and/or a derivative thereof may further comprise an additional active agent, wherein the compound and the additional active agent (s) are administered as a mixture, separately and simultaneously, or separately in any order.
  • additional active agent such as suramin and/or a derivative thereof
  • compositions comprising a compound of the disclosure are administered in combination with at least two additional active agents.
  • a composition comprising a compound disclosed herein can also be administered in combination with other classes of compounds, including, but not limited to, sepsis treatments, such as drotrecogin-a; steroidals, such as
  • hydrocortisone hydrocortisone
  • local or general anesthetics such as ketamine
  • platelet aggregation inhibitors such as clopidogrel
  • HMG-CoA reductase inhibitors such as atorvastatin
  • anticoagulants such as heparin
  • thrombolytics such as
  • sequestrants such as colestipol; non-steroidal anti-inflammatory agents (NSAIDs), such as naproxen; cholesteryl ester transfer protein (CETP) inhibitors, such as anacetrapib; anti-bacterial agents, such as ampicillin; anti-fungal agents, such as amorolfine; norepinephrine reuptake inhibitors (NRIs), such as atomoxetine; dopamine reuptake inhibitors (DARIs), such as methylphenidate ;
  • NSAIDs non-steroidal anti-inflammatory agents
  • CETP cholesteryl ester transfer protein
  • anti-bacterial agents such as ampicillin
  • anti-fungal agents such as amorolfine
  • norepinephrine reuptake inhibitors NRIs
  • DARIs dopamine reuptake inhibitors
  • sedatives such as diazepham; norepinephrine-dopamine reuptake inhibitor (NDRIs), such as bupropion; serotonin-norepinephrine- dopamine-reuptake-inhibitors (SNDRIs), such as venlafaxine;
  • NDRIs norepinephrine-dopamine reuptake inhibitor
  • SNDRIs serotonin-norepinephrine- dopamine-reuptake-inhibitors
  • monoamine oxidase inhibitors such as selegiline; hypothalamic phospholipids; endothelin converting enzyme (ECE) inhibitors, such as phosphoramidon; opioids, such as tramadol; thromboxane receptor antagonists, such as ifetroban; potassium channel openers; thrombin inhibitors, such as hirudin; hypothalamic phospholipids; growth factor inhibitors, such as modulators of PDGF activity; platelet activating factor (PAF) antagonists; anti-platelet agents, such as GPIIb/IIIa blockers (e.g., abdximab, eptifibatide, and tirofiban) , P2Y (AC) antagonists (e.g., clopidogrel, ticlopidine and CS-747), and aspirin; low molecular weight heparins, such as enoxaparin; Factor Vila Inhibitors and Factor Xa Inhibitors; renin inhibitors;
  • benzothiazide ethacrynic acid, tricrynafen, chlorthalidone, furosenilde, musolimine, bumetanide, triamterene, amiloride, and spironolactone
  • recombinant tPA streptokinase, urokinase, prourokinase , and anisoylated plasminogen streptokinase activator complex (APSAC)
  • anti-diabetic agents such as biguanides (e.g.
  • metformin glucosidase inhibitors
  • insulins e.g., acarbose
  • meglitinides e.g., repaglinide
  • sulfonylureas e.g., glimepiride, glyburide, and glipizide
  • thiozolidinediones e.g. troglitazone , rosiglitazone and pioglitazone
  • PPAR-gamma agonists e.g. troglitazone , rosiglitazone and pioglitazone
  • mineralocorticoid receptor antagonists such as spironolactone and eplerenone; growth hormone secretagogues ; aP2 inhibitors;
  • phosphodiesterase inhibitors such as PDE III inhibitors (e.g., cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil, vardenafil) ; protein tyrosine kinase inhibitors;
  • antiinflammatories such as methotrexate, FK506 (tacrolimus, Prograf) , mycophenolate mofetil; chemotherapeutic agents; immunosuppressants; anticancer agents and cytotoxic agents
  • alkylating agents such as nitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimines , and triazenes
  • antimetabolites such as folate antagonists, purine analogues, and pyridine analogues; antibiotics, such as anthracyclines ,
  • bleomycins bleomycins, mitomycin, dactinomycin, and plicamycin
  • enzymes such as L-asparaginase ; farnesyl-protein transferase inhibitors;
  • hormonal agents such as glucocorticoids (e.g., cortisone), estrogens /antiestrogens , androgens /antiandrogens , progestins, and luteinizing hormone-releasing hormone anatagonists , and octreotide acetate; microtubule-disruptor agents, such as ecteinascidins ;
  • microtubule-stabilizing agents such as pacitaxel, docetaxel, and epothilones A-F; plant-derived products, such as vinca alkaloids, epipodophyllotoxins , and taxanes; and topoisomerase inhibitors; prenyl-protein transferase inhibitors; and cyclosporins; steroids, such as prednisone and dexamethasone ; cytotoxic drugs, such as azathiprine and cyclophosphamide; TNF-alpha inhibitors, such as tenidap; anti-TNF antibodies or soluble TNF receptor, such as etanercept, rapamycin, and leflunomide; and cyclooxygenase-2 (COX- 2) inhibitors, such as celecoxib and rofecoxib; and miscellaneous agents such as, hydroxyurea, procarbazine, mitotane,
  • plant-derived products such as vinca alkaloids, epipod
  • complexes such as cisplatin, satraplatin, and carboplatin.
  • the disclosure also provides a methods of diagnosing mitochondrial diseases and disorders including chronic fatigue syndrome and autism spectrum disorders using various metabolic signatures or metabolomics information.
  • Metabolomics has several advantages over genomics for the diagnosis of complex chronic disease and for growing interest in precision medicine.
  • the genome represents an admixture of ancestral genotypes that were selected for fitness in ancestral environments.
  • the metabolic state of an individual at the time of illness is produced by both current conditions, age, and the aggregate history, timing, and magnitude of exposures to physical and emotional stress, trauma, diet, exercise, infections, and the microbiome recorded as metabolic memory.
  • metabolomics provides direct small molecule information
  • the results can provide immediately actionable treatment information using readily available small molecule nutrients, cofactors, and life style interventions.
  • the results presented herein demonstrates that chronic fatigue syndrome has an objectively identifiable chemical signature in both men and women and that targeted metabolomics can be used to uncover biological insights that may prove useful for both diagnosis, and personalized treatment.
  • German is an example of one well-studied system.
  • the developmental stage of dauer is a hypometabolic state capable of living
  • hypometabolic survival state that is triggered by environmental stress.
  • the metabolic features of chronic fatigue syndrome and dauer correspond to the same pathways that characterize the acute cell danger response and metabolic syndrome, but are regulated in the opposite direction.
  • cholesterol, phospholipids, and uric acid are often elevated in the acute cell danger response and metabolic syndrome, but these metabolites were decreased in chronic fatigue syndrome patients.
  • a prediction based on these findings is that patients with chronic fatigue syndrome would be more resistant to the constellation of hypertension, dyslipidemia, central obesity, and insulin resistance that increase all-cause mortality associated with metabolic syndrome, but at the cost of significant long-term disability, pain, and suffering.
  • NADPH NAD + dependent enzymes: 1) Malic enzyme (ME), 2) Isocitrate dehydrogenase (IDH), 3) Glutamate dehydrogenase (GDH) , 4) Nicotinamide nucleotide transhydrogenase (NNT) , and 5) Methylene tetrahydrofolate
  • MTHFD2L dehydrogenase
  • Each of these enzymes has at least one mitochondrial isoform and is known to be upregulated under conditions of environmental or developmental stress. It has recently been shown that mitochondrial MTHFD2L is responsible for producing 20-40% of cellular NADPH by the oxidation of methylene tetrahydrofolic acid to 10-formyl tetrahydrofolate . These data show that folates are not only important in methylation reactions, but also in regulating intracellular redox and NADPH levels (see FIG . 8 ) . A number of single nucleotide polymorphisms (SNPs) have been identified in the MTHFD2L gene that correlate with the cell danger response and ILlp production triggered by smallpox
  • NADPH nicotinamide nucleotide transhydrogenase
  • NADPH hydrogen peroxide
  • O 2 excess diatomic oxygen
  • NADPH When NADPH levels are higher, metabolism is shifted from persistence to normal cell function and growth, anabolic pathways are stimulated, biomass is created, and carbons and electrons are stored as biopolymers for cell growth and repair in the form of lipids, protein, glycogen, glycans, and nucleic acids.
  • NADPH is neither the problem nor the solution by itself. It is a messenger and cofactor. NADPH cannot work without the availability of hundreds of carbon skeletons of intermediary metabolism needed to carry out the message—the signal that fuel stores are either replete or limiting and metabolism must be adjusted accordingly. Specifically, NADPH cannot be simply added as a nutritional supplement to produce the tidal change in metabolism needed to shift the dauer state of chronic fatigue syndrome to normal health. Incremental improvements in NADPH production could theoretically be supported by
  • Chronic fatigue syndrome has a chemical signature that can be identified using targeted plasma metabolomics .
  • Receiver operator characteristic (ROC) curve analysis showed a diagnostic accuracy that exceeded 90%. The pattern and directionality of these changes showed that chronic fatigue syndrome is a conserved, hypometabolic response to environmental stress similar to dauer. Only about 25% of the metabolite disturbances found in each person were needed for the diagnosis of CFS . About 75% of the metabolite abnormalities were unique to the individual and useful in guiding personalized treatment. The finding of an objective chemical signature in chronic fatigue syndrome helps to remove diagnostic uncertainty, will help clinicians monitor individualized responses to treatment, and will facilitate multi-center clinical trials.
  • small molecule metabolite profile (s) or metabolomic profile (s) from a test subject or patient is/are compared to that/those of a control small molecule or control metabolomic profile.
  • detected amounts of metabolites are compared to normal or control amounts, such as amounts detected performing similar methods on a normal or control sample.
  • a normal or control sample in some aspects is one obtained from a subject who does not have, or is known not to have
  • Such comparisons can be made by individuals, e.g., visually, or can be made using software designed to make such comparisons, e.g., a software program may provide a secondary output which provides useful information to a user.
  • a software program can be used to confirm a profile or can be used to provide a readout when a comparison between profiles is not possible with a "naked eye".
  • the selection of an appropriate software program, e.g., a pattern recognition software program is within the ordinary skill of the art. An example of such a program is
  • test metabolite is intended to indicate a substance the concentration of which in a biological sample is to be measured; the test metabolite is a substance that is a by-product of or corresponds to a specific end product or intermediate of metabolism.
  • the collection of metabolomic data can be through, for example, a single technique or a combination of techniques for separating and/or identifying small molecules known in the art.
  • Small molecule metabolites can be detected in a variety of ways known to one of skill in the art, including the refractive index spectroscopy (RI), ultra-violet spectroscopy (UV) ,
  • NMR nuclear magnetic resonance
  • LS light scattering analysis
  • HPLC high pressure liquid chromatography
  • MALDI-TOF matrix-assisted laser desorption ionization-time of flight
  • a biological sample obtained from a subject can be prepared for use in one or more of the foregoing identification/detection methods.
  • the biological sample can be divided for multiple parallel measurements and/or can be enriched for a particularly type of small molecule metabolite ( s ) .
  • different fractionation procedures can be used to enrich the fractions for small molecules.
  • small molecules obtained can be passed over several fractionation columns. The fractionation columns will employ a variety of detectors used in tandem or parallel to generate the small molecule metabolite profile.
  • the biological sample will be fractionated on HPLC columns with a water soluble array.
  • the water soluble small molecule metabolites can then be detected using fluorescence or UV detectors to generate the small molecule metabolite profiles.
  • hydrophobic columns can also be used to generate small molecule metabolite profiles.
  • CFS chronic fatigue syndrome
  • mitochondrial related disease e.g., chronic fatigue syndrome (CFS) and/or mitochondrial related disease by analyzing metabolites found in easily obtained biospecimens (e.g., blood, urine) .
  • the methods of the disclosure allows clinicians to stratify subjects according to the risk of CFS or the occurrence of CFS.
  • the methods use high performance liquid chromatography (HPLC) chromatography, tandem Mass Spectrometry (LC- MS/MS) , and analytical statistical techniques to identify and analyze metabolomic profiles.
  • HPLC high performance liquid chromatography
  • MS/MS tandem Mass Spectrometry
  • the methods of the disclosure can utilize the measurement of a thousand or more metabolites (e.g., up to 2500 or more) or fewer than 2500 (e.g., 15-30, 30-60, 60-100, 100-200, 200-500, 500- 1000, 1000-1500, 1500-2000, 2000-2500 and any number there between 15 and 2500) . While several hundred small molecule metabolites can be measured, in practice 30 or fewer small molecule metabolites may be sufficient for diagnostic and prognostic purposes. Furthermore, the small molecule metabolites being measured can include more than one metabolite from a particular metabolic pathway. Thus, for example, 30 or fewer small molecule metabolites may be
  • small molecule metabolites are collected and subjected to chemical extraction. Internal isotopically labeled standards can be added to the sample and injected into an HPLC-Mass Spectrometer. Small molecule metabolites are separated and then measured via mass spectrometry. Subjects having or at risk of having CFS (or other mitochondrial disease or disorder to be analyzed) have a distinct set of metabolites (e.g., a "CFS small molecule metabolite profile”) that are indicative of a CFS metabolomic profile that distinguish them from healthy controls.
  • the small molecule metabolites are collected, processed to non-naturally occurring analytes (e.g., mass fragments), the analytes processed to determine their identities and the data plotted in 2D or 3D coordinates and compared to a control small molecule metabolite profile or a control metabolomics profile, which can be plotted on the same coordinate system (e.g., a mass spectroscopy plot, an HPLC plot or the like) . This plot can then be output to a user or medical technician for analysis.
  • analytes e.g., mass fragments
  • the method of the disclosure includes obtaining a small molecule metabolite profile from a test subject, identifying small molecule analytes that are over produced or under produced (including presence and absence) generating a metabolomics profile which is indicative of the activity of the various metabolic pathways associated with the small molecule metabolites and comparing metabolomics profiles of the test subject/patients to a standard, normal control metabolomics profile.
  • an over or under production of a metabolite compared to a control by at least 2 standard deviations is indicative of an aberrant metabolic pathway.
  • a difference in the amount of metabolite by 10% or more (e.g., 10%-100% or more) compared to a control value is indicative of an aberrant metabolic pathway.
  • the method thus involves identifying the small molecules which are present in aberrant amounts in the test small molecule metabolite profile.
  • the small molecules present in aberrant amounts are indicative of a diseased or dysfunctional metabolic pathway.
  • An "aberrant amount” includes any level, amount, or concentration of a small molecule metabolite, which is different from the level of the small molecule of a standard sample by at least 1 standard deviation (typically 2 standard deviations are used) .
  • the aberrant amount can be higher or lower than the control amount.
  • the method of the disclosure includes measuring a plurality of pathways and metabolites.
  • Tables 2-4 provides an exemplary list of such pathways and an exemplary number of metabolites that can be measure in each pathway.
  • a method comprises obtaining a sample from a subject (e.g., blood, urine, tissue); preparing the sample (e.g., extracting, enriching, and the like) metabolites, which can include the addition of internal standards; performing a technique to quantitate metabolites in the sample (e.g., HPLC, Mass
  • CFS metabolomics profile 5 to 61 metabolites were determined in one study to be useful in characterizing a CFS subject (see, e.g., Tables 11 and 12) .
  • the number of metabolites that can be used to characterize a CFS subject is selected from 7, 10, 12, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or a range between any two of the foregoing numbers. It should be noted herein, that CFS metabolomics profiles can be further differentiated based upon the sex of the subject, wherein the optimal metabolomics profile that is
  • the disclosure provides methods of using metabolomics profile information to study the effectiveness of a therapy or intervention for CFS, such as the administration of antipurinergic compounds like suramin and/or a derivative thereof to a subject. For example, by obtaining and comparing the metabolomics profiles, amounts of metabolites, and/or alterations in pathways, from a subject having CFS and a control population, certain aberrant small molecule metabolites can be identified and their corresponding metabolic pathways identified.
  • a therapy or intervention for CFS such as the administration of antipurinergic compounds like suramin and/or a derivative thereof to a subject.
  • antipurinergic compounds can then be administered or provided to a subject having CFS and a small molecule metabolite profile and metabolomics profile obtain from the subject during or after therapy.
  • the small molecule and metabolomics profiles from the subject are analyzed with particular attention to any
  • a change in the small molecule metabolite or metabolomics profile of the treated subject that is more consistent with a normal control profile would be indicative of an effective therapy.
  • more consistent means that the aberrant values or pathway are trending towards or are within a desired range considered “normal” for the population.
  • antipurinergic therapy e.g., administration of suramin
  • APT antipurinergic therapy
  • antipurinergic therapy is ultimately traceable to mitochondria which is regulated by purinergic signaling.
  • Treatment with antipurinergic compounds is expected to turn off the cell danger response (CDR) which is controlled by purinergic signaling, which has been identified herein as a component of chronic fatigue syndrome.
  • CDR cell danger response
  • the primary pharmacologic mechanism of action of antipurinergic compounds like suramin, is as a competitive antagonist of extracellular ATP and other nucleotides, acting at purinergic receptors.
  • antipurinergic agents that block P2X/P2Y signaling or extracellular ATP, ADP, and/or Pl-receptor (Adenosine) signaling by extracellular adenosine or AMP.
  • antipurinergic compounds that can be used in the methods disclosed herein, include but are not limited to, P2Y inhibitors like 2-methylthioladenosine monophosphate, A3P5PS, AMPaS,
  • diadenosine tetraphosphate AR-C66096, AR-C67085MX, AR-C69931MX, AR-C118925XX, C1330-7, Cangrelor, Clopidrogel, Elinogrel, IP51, MRS-2179, MRS-2211, MRS-2279, MRS-2395, MRS-2500, MRS-2578, NF-157, NF-340, 2 , 2 ' -pyridylisatogen tosylate, pyridoxalphosphate- 6- azophenyl-2 ' , 4 ' -disulfonic acid, prasugrel, PSB-0739, RB-2, regrelor, suramin, ticagrelor, and ticlopidine.
  • LC-MS/MS analysis was performed by scheduled multiple reaction monitoring (sMRM) under Analyst vl.6.2 software control in both negative and positive mode with rapid polarity switching (50 ms) . Nitrogen was used for curtain gas (set to 30), collision gas
  • ion source gas 1 and 2 (set to 35) .
  • the source temperature was 500 °C.
  • Spray voltage was set to -4500 V in negative mode and 5500 V in positive mode.
  • the values for Ql and Q3 mass-to-charge ratios (m/z) , declustering potential (DP) , entrance potential (EP) , collision energy (CE) , and collision cell exit potential (CXP) were determined and optimized for each MRM for each metabolite.
  • the network for Chronic Fatigue Syndrome included the 20 metabolic pathways associated with the cell danger response (36) that were dysregulated.
  • Nodes in the Cytoscape network represent metabolites within the pathways and have been colored according to the Z-score.
  • the Z-score was computed as the arithmetic difference between the mean concentration of each metabolite in Chronic Fatigue and controls, divided by the standard deviation in the controls. Node colors were arranged on a red-green color scale with green
  • the average age of men with CFS in this study was 53 (+/- 2.8) (see TABLE 7) .
  • the average age of the women with CFS was 52 (+/- 2.5) .
  • the average age of onset was 30 (+/- 2.6) years for the men and 33 (+/- 2.3) years for the women.
  • the average duration of illness was 21 (+/- 3.0) years for men and 17 (+/- 2.3) years for women.
  • the Karnofsky quality of life performance score for men was 62 (+/- 3.2), and 54 (+/- 3.3) for women (see Table 7).
  • Plasma adipoylcarnitine is a six-carbon dicarboxylic acid (CeDC) carnitine ester that was increased in females with chronic fatigue syndrome, but not in males (see TABLES 8-9) .
  • Elevations in adipoylcarnitine are sensitive indicator of decrease in riboflavin dependent mitochondrial beta oxidation of fatty acid, and fasting. Fasting, or decreased intracellular allocation of fatty acids for oxidation in mitochondria results in the induction of the peroxisomal enzyme known as the NAD +
  • L-bifunctional enzyme activated, L-bifunctional enzyme (enoyl-CoA, hydratase/3- hydroxyacyl CoA dehydrogenase, EHHADH) .
  • This enzyme is required for the synthesis of medium chain dicarboxylic acids like adipic acid.
  • Spurious elevations of adipoylcarnitine can be seen as the result of high volume consumption of adipic acid-containing gelatin.
  • Plasma 2-arachidinoyl glycerol (2AG) was decreased in females with CFS, but not in males (see TABLES 8-9) .
  • 2AG is a natural cannabinoid agonist of CBi signaling.
  • CBi receptors Inhibition of CBi receptors is known to inhibit fatty acid oxidation and increase plasma levels of adipoyl carnitine.
  • the observed decrease in 2AG might contribute to decreased oxidation of adipic acid and increased plasma adipoylcarnitine.
  • Plasma 2-octenoylcarnitine is an eight-carbon mono- unsaturated fatty acid (C 8 :1) carnitine ester that was decreased in females with chronic fatigue syndrome, but not in males (see TABLES 8-9) . Elevations of octenoylcarnitine are seen in calorie excess conditions such as obesity and metabolic syndrome. Decreased octenoic acid is also known to inhibit the susceptibility to Herpes virus infections and to increase cell membrane stiffness. Octenoic acid is produced as an intermediate of mitochondrial fatty acid and lipoic acid synthesis.
  • acylcarnitine It is the substrate for mitochondrial enoyl thioester reductase (ETR) , a family of enzymes that requires NADPH to reduce the double bond to octanoic acid, which is then used for lipoic acid synthesis. Decreased levels of this Cs : 1 acylcarnitine are consistent with decreased mitochondrial fatty acid synthesis, increased oxidation, increased renal secretion, or a combination of the three. All other acylcarnitine species measured (C 2 -C24) were normal in both males and females. No abnormalities of fatty acid oxidation were found in males with chronic fatigue syndrome.
  • Serine and Threonine were Increased in Males. Plasma serine and threonine levels were increased in males with chronic fatigue syndrome, but not in females (see TABLES 8-9) .
  • the intracellular ratio of serine to glycine is a key regulator of 1- carbon, nucleotide, and folate metabolism.
  • L-Serine is also an essential precursor of sphingolipid, phosphatidylserine , D-serine, and de novo cysteine and glutathioine synthesis. Both serine and threonine are gluconeogenic amino acids that can be used to make glucose during fasting or stress. Acute stress or infection results in a decrease in both plasma serine and threonine levels.
  • sphingolipids are a recently recognized hallmark of obesity, metabolic syndrome, insulin resistance, and a risk factor in
  • Alzheimer dementia These abnormalities are improved by exercise, and caloric restriction. Effective methods to increase abnormally low sphingolipids have not yet been developed.
  • desmosterol accumulates in cell membranes, but does not change significantly in plasma.
  • Membrane accumulation of desmosterol inhibits endocytosis and pinocytotic import through caveolae, and inhibiting the production of sphingolipid-rich and cholesterol-rich lipid rafts needed for cell-cell signaling. This would serve as a cell defense mechanism that inhibits the uptake of certain intracellular bacterial pathogens as Coxiella (Q fever) and Borrelia (Lyme disease) .
  • Metabolomic data were log-transformed, scaled by control standard deviations, and analyzed by multivariate partial least squares discriminant analysis (PLSDA) , principal components analysis (PCA) , t test, univariate ANOVA with pairwise comparisons and post hoc correction for multiple hypothesis testing using Fisher's least significant difference method in MetaboAnalyst, or the false discovery rate (FDR) method of PLSDA and PCA.
  • PLSDA multivariate partial least squares discriminant analysis
  • PCA principal components analysis
  • FDR false discovery rate
  • AUROC AUROC curve analysis. Classifier robustness was estimated by repeated double cross validation (rdCV) , and permutation testing 1000 times in MetaboAnalyst. Confidence intervals for the ROC curves were calculated by bootstrap resampling. Sensitivity, specificity, accuracy, positive predictive value, negative predictive value, and number of misclassifications were estimated by conventional 2 x 2 contingency table analysis and p values calculated by Fisher's exact test in Prism.
  • Chronic Fatigue Syndrome Patients had an Average of 40 Metabolic Abnormalities.
  • the mean number of metabolites falling outside of the 95% confidence limits (beyond 2 standard deviations from the mean) in females with chronic fatigue patients was 39 (+/- 3.5) (mean +/- SEM) , and 41 (+/- 4.4) in males (See FIG 3A-B) . This was significantly more than the number found among age and sex- matched controls, which was 16 (+/- 1.8) (p ⁇ 2 x 10 -6 ; see FIG 3A- B) .
  • the mean and 95% confidence interval for the number of abnormalities expected by chance in the controls was calculated from the binomial distribution of 420 measured metabolites with a mean chance of abnormality equal to the p value of 0.05 for each test.
  • the metabolites that were altered most in chronic fatigue syndrome were ranked by multivariate analysis (see FIG. 4A-B) . Individual metabolite abnormalities were visualized according to a
  • sphingolipids are synthesized from the amino acid serine and palmitoyl-CoA. Ceramides are produced rapidly from sphingomyelins by the inducible enzyme acid sphingomyelinase (ASMase) and redox regulated neutral sphingomyelinase (nSMase) . Ceramides, and gangliosides made from ceramides, are important components of cell membrane patches and microdomains called lipid rafts that mobilize and aggregate during cell activation or infection. Sphingolipid rafts are enriched in cholesterol, and are crucial for cell-cell signaling, and reactive oxygen species (ROS) production for oxidative shielding and defense. Ceramide-, ganglioside- , and cholesterol-enriched cell membrane rafts are needed to recruit protein receptor and effector subunits during cell activation, cardiovascular and renal disease, and bacterial and protozoal infections. Increased membrane expression and release of
  • sphingolipids is a universal response to acute infection, but can also be hijacked as a vehicle for virus entry and spread, and can result in autoimmune responses to gangliosides, and to lipid raft- associated receptors like CD46. Down-regulation of sphingolipids in chronic fatigue syndrome is consistent with a compensatory response to environmental stress, inflammation, or infection.
  • Sphingolipids and Glycosphingolipids were Decreased. The largest disturbances in the chemical signature of chronic fatigue syndrome were produced by widespread decrease in plasma sphingo- and glycosphingolipids (see FIG. 2C-D, see Tables 8-9) . Thirty molecular species of sphingolipids were decreased in males, and 21 were decreased in females. Sphingolipid and glycosphingolipid abnormalities explained 55% of the metabolic impact in males, and 44% in females (Tables 8-9) . Measured glycosphingolipids included glucosyl- (GC) , dihexosyl- (DHC) , and trihexosyl (THC) ceramides.
  • GC glucosyl-
  • DHC dihexosyl-
  • THC trihexosyl
  • sphingomyelins in females see Tables 1-4.
  • females with chronic fatigue retained more sphingomyelin species in the normal range than males.
  • the low sphingolipid profile in chronic fatigue syndrome appears to be an adaptive response that is opposite to the increased sphingolipids observed in metabolic syndrome and the acute cell danger response, and ultimately may represent a fundamental response to oppose the spread of persistent viral and intracellular bacterial infections.
  • PC phosphatidylcholine
  • Plasma uric acid was decreased in males with chronic fatigue syndrome (see Table 8) .
  • Uric acid is the end product of purine metabolism and an important antioxidant molecule.
  • Plasma adenosine was decreased in females (see Table 9) .
  • Plasma adenosine is produced from ATP and ADP released from cell surface ectonucleotidases , and by S-adenosylhomocysteine hydrolase
  • SAHH chronic fatigue syndrome
  • Aromatic Amino Acid Metabolites from the Microbiome were Decreased.
  • Plasma 4-hydroxyphenyllactic acid (HPLA) was decreased in males with chronic fatigue syndrome (see Table 8) .
  • Plasma phenyllactic acid (PLA) was decreased in females (see Table 9) .
  • HPLA is a microbiome metabolite of tyrosine.
  • PLA is a microbiome metabolite of phenylalanine. This pattern is also opposite of what is found during acute inflammation, and infection.
  • Flavin Adenine Dinucleotide was Decreased. Plasma FAD was decreased in both males and females with chronic fatigue syndrome (see Tables 8- 9 ) . FAD is synthesized from riboflavin
  • vitamin B2 vitamin B2
  • ATP ribulose-1-phosphate
  • GI absorption, distribution, and transporter-mediated uptake of FAD are carefully regulated during health and disease.
  • FAD is mobilized from tissues, increased in the plasma, and renal secretion is increased under conditions of stress or infection.
  • FAD is an important cofactor for fatty acid oxidation and sterol synthesis, is required for activation and oxidation of vitamin B6 (pyridoxine) , lipoic acid metabolism (E3 subunit) needed for pyruvate, alpha-ketoglutarate, and branched chain amino acid oxidation, vitamin A activation, 5- methyltetrahydrofolic acid synthesis, niacin and NAD synthesis, and glutathione reduction.
  • vitamin B6 pyridoxine
  • E3 subunit lipoic acid metabolism
  • riboflavin deficiency can be produced by dietary and environmental factors. Severe riboflavin deficiency can present with a plasma acyl-carnitine pattern similar to multiple acyl-CoA dehydrogenase deficiency (MADD) , also known as glutaric aciduria type II (GAII) . GAII-like acyl-carnitine abnormalities did not appear in chronic fatigue syndrome patients.
  • MADD multiple acyl-CoA dehydrogenase deficiency
  • GAII glutaric aciduria type II
  • the Bloch pathway is normally used preferentially in certain metabolic stress-response tissues like the gonads, spleen, adrenal glands, kidney, and adipose tissue.
  • the liver uses a nearly-equal blend of Bloch and K-R pathways.
  • the data are consistent with increased flux through the desmosterol pathway to maintain normal cellular levels of cholesterol.
  • the desmosterol pathway corresponds to the stress-inducible arm of de novo cholesterol and sterol synthesis.
  • CDCA Plasma chenodeoxycholic acid
  • P5C Pyrroline-5-carboxylic acid
  • P5C production is a well-studied response to stress in plants, and mammals.
  • P5C can be produced by the stress-induced oxidation of proline and hydroxyproline from collagen turnover via the enzyme proline oxidase, or from glutamate oxidation via pyrroline-5- carboxylate synthase (P5CS) .
  • P5C is converted to glutamate semialdehyde (GSA) non-enzymatically, then to ornithine under stress conditions.
  • GSA glutamate semialdehyde
  • hydroxyproline is glyoxylate, which can be transaminated in mitochondria to produce glycine, and metabolized in peroxisomes ⁇ oxalate and peroxide for cell defense, innate and antiviral immunity .
  • Arginine is both a source of urea by arginase in the urea cycle, but more importantly, it is an activator of iV-acetylglutamate (NAG) synthesis.
  • NAG is the obligate activator of carbamoyl phosphate synthetase I (CPS-I) .
  • CPS-I is required for the introduction of ammonia into the urea cycle via the synthesis of citrulline from ornithine and
  • Citrulline, ornithine, proline, glutamine, and glutamate levels were all normal. Under stress conditions, proline from collagen breakdown is shunted to arginine synthesis to spare nitrogen from other amino acids, and limit wasting during periods of decreased calorie and or protein intake. Increased arginine might
  • ADMA asymmetric dimethylarginine
  • HICA 2-Hydoxyisocaproic acid
  • flavin adenine dinucleotide FAD
  • pyroglutamic acid also known as 5-oxoproline
  • HICA 2-hydroxyisocaproic acid
  • L-serine L-serine
  • lathosterol L-serine
  • bl-analyte phosphatidyl choline PC ( 16 : 0/16 : 0 )
  • ceramide (dl 8 : 1 /22 : 2 ) , lathosterol, adenosine, phosphatidylinositol PI (16: 0/16: 0) , flavin adenine dinucleotide (FAD), 2- octenoylcarnitine , phosphatidyl choline plasmalogen PC (22 : 6/P18 : 0) , phosphatidyl choline PC (18 : 1/22 : 6) , l-pyrroline-5-carboxylate (P5C) , and chenodeoxycholic acid (CDCA) .
  • dAUROC area under the receiver operator curve reflects the overall accuracy of diagnosis using these analytes.
  • erdCV repeated random sub-sample (2/3 in, 1/3 out) double cross validation .
  • fPermutation p values represent the probability that the random forest classification of cases and controls using the specified analytes could be obtained by chance.
  • PCA principal components analysis
  • ADOS-2 ADOS-2
  • EOWPVT Expressive One Word Picture Vocabulary Testing
  • CGI Clinical Global Impression
  • CGI Clinical Global Impression
  • a 24-question Clinical Global Impression (CGI) instrument was developed to assess the core symptoms of autism spectrum disorders and some of the most common comorbid features.
  • the CGI instrument scoring system was the traditional 7-point, CGI-Improvement scale. In this scale, the historian gives a score of 0 if the symptom "was never a problem", a 1 for "very much improved", a 4 for "no change", and a 7 for "very much worse”.
  • the parents were asked to write in the top 3 symptoms or behaviors that were most changed over the 6 weeks since the suramin infusion.
  • Randomization and Masking 20 males with ASD were screened. 16 met entry criteria. 10 participants could be matched by age, non-verbal IQ, and ADOS scores into 5 pairs. The
  • randomization sequence was generated electronically by the biostatistical team. Subjects within each pair were allocated to receive suramin or saline according to the prospectively determined randomization sequence. The randomization sequence was concealed from the clinical team and implemented by the investigational pharmacy, which prepared drug and placebo for infusion. The design was double-blind. The mask was not broken until all subjects had completed the study and all clinical data had been collected.
  • Storyboards and accompanying social stories were created to illustrate each step of the study for parents to review with each child before the study.
  • Protocol Deviations The original protocol was designed to collect electroencephalography (EEG) , heart rate variability (HRV) , balance, gait, fine motor, and sensory motor data as secondary outcomes.
  • EEG electroencephalography
  • HRV heart rate variability
  • HRV heart rate variability
  • balance gait
  • fine motor fine motor
  • sensory motor data as secondary outcomes.
  • PPVT Peabody Picture Vocabulary testing
  • Suramin was provided as the hexasodium salt (MW 1429.2 g/mol) in 1 gram lyophilized vials by Bayer Pharmaceuticals, Inc. A 1-gram vial was reconstituted in 10 mL of sterile water for infusion to prepare a 10% (100 mg/mL) solution. Height and weight were recorded, vital signs and capillary oxygen saturation (pulse oximetry) measured, physical and neurological examinations were conducted, and urine and blood for safety monitoring, pharmacology, and metabolomics was collected before the infusion. Each child then received a 50 mg test dose
  • Safety and Adverse Event Monitoring Blood and urine for safety and toxicity monitoring were collected immediately before the infusion, 1 hour after the infusion, 2 days after, and 45 days after the infusion. Vital signs and anthropomorphic measurements were also collected. Safety surveillance included 18 vital sign and anthropometric features, 19 complete blood count (CBC) parameters, 20 blood chemistry measures, 3 thyroid and Cortisol measures, and 5 lipid measures at the 5 time points. 24 urinalysis features were measured at 4 times: baseline, pre-infusion, 2-days post-infusion, and 45-days post-infusion.
  • CBC complete blood count
  • PK suramin pharmacokinetics
  • LC-MS/MS tandem mass spectrometry
  • PK parameters were scaled allometrically with volume terms scaled to linear body weight (kgl.O) and clearance terms scaled to weight (kg0.75) .
  • Scaled adult suramin parameters of compartmental volumes of distribution and clearance were used as initial parameter estimates and between subject variability only estimated for clearance (CL) and the peripheral volume of distribution (Vd) .
  • FIG. 9 illustrates the CONSORT flow diagram for patient recruitment, allocation, and analysis in the SAT-1 study.
  • the two treatment groups were well matched (Table 14) .
  • the distribution phase half-life was 7.4 ⁇ 0.55 hours.
  • the suramin levels rapidly fell below 100 ⁇ and into the target range before day 2 in all subjects, with an average plasma level of suramin of 12.0 ⁇ 1.5 ⁇ on day 2 (see FIG. 12B, and Table 16) .
  • Target concentrations of 1.5-15 ⁇ were maintained between 2 days and 6 weeks following the dose (see FIG. 12) .
  • the steady-state volume of distribution was 0.83 ⁇ 0.014 L/kg (22.7 ⁇ 2.6 L/m 2 ) .
  • the clearance was 1.95 ⁇ 0.21 mL/hr/kg (0.056 ⁇ 0.011 L/hr/m 2 ) .
  • the terminal elimination phase half-life (tl/2) was 14.7 ⁇ 1.4 days
  • the resulting fresh lithium-heparin plasma was transferred to labeled 1.2 mL or 2.0 mL externally threaded, cryotubes with a minimum headspace air gap for storage at -80 °C for analysis.
  • Samples were analyzed on an AB SCIEX QTRAP 5500 triple quadrupole mass spectrometer equipped with a Turbo V electrospray ionization (ESI) source, Shimadzu LC-20A UHPLC system, and a PAL CTC autosampler.
  • ESI Turbo V electrospray ionization
  • 90 ⁇ of plasma was thawed on ice and transferred to a 1.7 mL Eppendorf tube.
  • Macromolecules protein, DNA, RNA, glycans, etc. were precipitated by extraction with 4 volumes (400 ⁇ L) of cold (-20 °C) ,
  • acetonitrile :methanol 50:50 (LCMS grade, Cat# LC015-2.5 and GC230-4, Burdick & Jackson, Honeywell), vortexed vigorously, and incubated on crushed ice for 10 min, then removed by centrifugation at 16,000g x 10 min at 4°C.
  • the supernatants containing the extracted metabolites and internal standards in the resulting 40:40:20 solvent mix of acetonitrile : methanol : water were
  • LC-MS/MS analysis was performed by scheduled multiple reaction monitoring (sMRM) under Analyst vl.6.2 software control in both negative and positive mode with rapid polarity switching (50 ms) . Nitrogen was used for curtain gas (set to 30), collision gas
  • ion source gas 1 and 2 (set to 35) .
  • the source temperature was 500 °C.
  • Spray voltage was set to -4500 V in negative mode and 5500 V in positive mode.
  • the values for Ql and Q3 mass-to-charge ratios (m/z) , de-clustering potential (DP) , entrance potential (EP) , collision energy (CE) , and collision cell exit potential (CXP) were determined and optimized for each MRM for each metabolite.
  • Targeted plasma metabolomics was performed immediately before infusion, at 2 days, and 6 weeks after the infusion.
  • the rank order of the top 35 of 48 significant metabolites 6-weeks after suramin treatment is illustrated in FIG. 13A.
  • the rank order after 2-days is illustrated in FIG. 14. Consistent with our previously published work using mouse models, the metabolic effects of suramin resulted in a decrease of the cell danger response and restored more normal metabolism (Table 17) .
  • FIG. 13B illustrates the similarities found in the pharmacometabolomic response to suramin in MIA13 and Fragile X mouse models and in children with ASD in this study. Twenty-one of the 28 pathways
  • Suramin Quantitation Suramin Quantitation. Suramin concentrations were measured by LC-MS/MS as previously described with modifications. Plasma suramin samples were collected at 1 hour, 2 days and 42 days post- infusion. Heparinized plasma, 90 ⁇ was used. Ten (10) ⁇ L of 50 ⁇ stock of trypan blue was added to achieve an internal standard concentration of 5 ⁇ . This was incubated at room temperature for 10 min to permit metabolite interaction with binding proteins, then extracted with 4 volumes (400 ⁇ of pre-chilled methanol- acetonitrile (50:50) to produce a final concentration of 40:40:20
  • Absolute concentrations of suramin were determined using a standard curve prepared in plasma to account for matrix effects, and the peak area ratio of suramin to the internal standard trypan blue.
  • the declustering potential (DP) , collision energy (CE) , entrance potential (EP) and collision exit potential (CXP) were -104, -9.5, -32 and -16.9, and -144.58, -7, -57.8 and -20.94, for suramin and trypan blue, respectively.
  • the ESI source parameters were set as follows: source temperature 500 °C; curtain gas 30; ion source gas 1, 35; ion source gas 2 35; spray voltage -4500 V. Analyst vl .6 was used for data acquisition and analysis.
  • ADOS-2 and expressive one-word picture vocabulary (EOWPVT) scores Table 21
  • Parents reported that after suramin treatment, the rate of language, social, behavioral, and developmental improvements continued to increase for 3 weeks, then gradually decreased toward baseline over the next 3 weeks.
  • the blood levels of suramin at 3 weeks were estimated to be 4.2 ⁇ 0.5 ⁇ using our PK model.
  • ADOS comparison scores were improved in the suramin treatment group at 6-weeks (see FIG. 16A-B) but were unchanged in the saline group (See FIG. 16A, and 16F) .
  • ADOS scores at 2-days after treatment were not changed (See FIG. 16E) .
  • EOWPVT scores were not changed (See FIG. 161) .
  • Secondary outcomes included Aberrant Behavior Checklist (ABC) , Autism Treatment Evaluation Checklist
  • Suramin has objective central nervous system (CNS) effects in animal models and children with autism despite being unable to penetrate the blood brain barrier. Suramin also has a number of peripheral effects on innate immunity, metabolism, pain, gut, autonomic, inflammatory, and other pathways regulated by purinergic signaling that may contribute to the beneficial effects observed. Previous studies have shown that suramin is taken up into the CNS at the level of the brainstem, although not appreciably into the cerebrum or cerebellum. There are 8 circumventricular organs (CVOs) in the brain that contain neurons that lack a blood brain barrier.
  • CVOs circumventricular organs
  • the area postrema in the brainstem is one of these CVOs that monitors the chemistry of the blood and transduces this information to higher centers in the brain for neuroendocrine, affective, cognitive, and behavioral integration.
  • the peripheral actions and indirect CNS effects of suramin may have certain advantages by minimizing the risk of CNS toxicity. While new antipurinergic drugs (APDs) may soon be developed that can pass the blood brain barrier, this appears not to be required to produce the behavioral effects of suramin in ASD.
  • the SAT-1 trial examined the effects of low-dose suramin or placebo in 10 children with autism spectrum disorder. No safety concerns were found.
  • a two-compartment pharmacokinetic model permitted accurate forecasting of plasma drug levels from 1 hour to 6-weeks after the infusion.
  • Metabolomic studies confirmed the importance of the cell danger response (CDR) 8 and purinergic signaling.
  • CDR cell danger response
  • a single, 20 mg/kg intravenous dose of suramin was associated with improved scores for language, social interaction, and decreased restricted or repetitive behaviors measured by ADOS, ABC, ATEC, and CGI scores. None of these improvements occurred in the 5 children who received placebo.
  • a rotarod test was used to detect difference between treatment groups.
  • a decrease in rotarod latency to fall in the Poly ( IC) -exposed animals was not shown.
  • antipurinergic therapy with suramin showed a trend toward improved rotarod performance in the females
  • Hyperactivity It was found that hyperactivity, measured by center entries in a standard open field test, was sexually dimorphic in this cohort of ASD mice. Male ASD mice showed an increased number (see FIG. 19) of center entries. This increase was returned to control levels by antipurinergic therapy with suramin. Females did not show hyperactivity and did not show a suramin effect (data not shown) .
  • a Single Dose of Suramin Corrects the Short-term Memory Defect in the Poly(IC) Mouse Five month old, ASD-like animals displayed a short-term memory defect that was quantified using the T-maze paradigm. Control C57BL/6 mice will recall the immediately previous T-maze arm exploration and will spontaneously alternate into right and left arms 75% of the time (see FIG. 23; Sal-Sal) . This alternation falls to random choice (50%) in ASD-like animals
  • antipurinergic therapy was sometimes more effective in males, as in the social approach paradigm and hyperactivity (see FIG. 17 and FIG. 19) . In some cases, it was more effective in females, as in correcting the coordination abnormalities (see FIG. 18) . In many cases,
  • antipurinergic therapy with suramin was equally effective in both males and females, as seen in the SIH and in the restoration of normal core body temperatures (see FIG. 20 and FIG. 21) .
  • Antipurinergic therapy holds great promise in treating this fundamental defect in autism.
  • Antipurinergic therapy is also effective in adult animals, 6 months of age (see FIG. 23 and FIG. 24) . In these animals, a single dose of suramin corrected behavioral and learning abnormalities that were present since shortly after birth. The results suggest that APT is capable of triggering a previously unknown, neurochemical switch that lies at the cause of abnormalities that were formerly thought to be a fixed feature of disease.
  • APT will be effective for at least treating the following: autism, chronic fatigue syndrome, fibromyalgia, obsessive compulsive disorder, generalized anxiety disorder, schizophrenia, bipolar depression, subacute therapy for traumatic brain injury (TBI), post-traumatic stress disorder (PTSD) , chronic traumatic encephalopathy (CTE) , disabling Attention Deficit
  • ADHD Hyperactivity Disorder

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Abstract

L'invention concerne des biomarqueurs utiles pour diagnostiquer et prédire le développement du syndrome de fatigue chronique (SFC). Elle concerne en outre des méthodes pour rétablir le métabolisme et faciliter la guérison chez des patients atteints de SFC par administration de composés anti-purinergiques.
PCT/US2017/041932 2016-07-14 2017-07-13 Diagnostic et méthodes de traitement du syndrome de fatigue chronique et des troubles du spectre autistique WO2018013811A1 (fr)

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020160108A1 (fr) * 2019-01-30 2020-08-06 Arizona Board Of Regents On Behalf Of The University Of Arizona Biomarqueurs lipidiques pour le dépistage et la surveillance du cancer
WO2020247127A1 (fr) * 2019-06-07 2020-12-10 Paxmedica, Inc. Compositions et méthodes pour traiter des troubles du système nerveux central
US11026904B2 (en) 2019-01-28 2021-06-08 Mitochondria Emotion, Inc. Mitofusin activators and methods of use thereof
US11083699B2 (en) 2019-01-28 2021-08-10 Mitochondria Emotion, Inc. Trans-4-hydroxycyclohexyl phenyl amide mitofusin activators and methods of use thereof
CN114839298A (zh) * 2019-12-28 2022-08-02 中精普康(北京)医药科技有限公司 一种用于检测结直肠癌或腺瘤的生物标志物及其方法
JP7360807B2 (ja) 2019-04-26 2023-10-13 株式会社東レリサーチセンター 疲労状態の検出を補助する方法
CN114839298B (zh) * 2019-12-28 2024-06-07 中精普康(北京)医药科技有限公司 一种用于检测结直肠癌或腺瘤的生物标志物及其方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130237454A1 (en) * 2010-05-12 2013-09-12 Steven E. Schutzer Diagnostic markers for neuropsychiatric disease
US20130267441A1 (en) * 2010-12-16 2013-10-10 Autism Biotech Limited Novel biomarker and uses thereof in diagnosis, treatment of autism
WO2015027116A1 (fr) * 2013-08-21 2015-02-26 The Regents Of The University Of California Motifs de métabolites pour le diagnostic et la prédiction de troubles affectant le cerveau et le système nerveux
US20150125526A1 (en) * 2013-11-06 2015-05-07 Raptor Pharmaceuticals Inc. Use of Cysteamine and Derivatives Thereof to Treat Mitochondrial Diseases

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130237454A1 (en) * 2010-05-12 2013-09-12 Steven E. Schutzer Diagnostic markers for neuropsychiatric disease
US20130267441A1 (en) * 2010-12-16 2013-10-10 Autism Biotech Limited Novel biomarker and uses thereof in diagnosis, treatment of autism
WO2015027116A1 (fr) * 2013-08-21 2015-02-26 The Regents Of The University Of California Motifs de métabolites pour le diagnostic et la prédiction de troubles affectant le cerveau et le système nerveux
US20150125526A1 (en) * 2013-11-06 2015-05-07 Raptor Pharmaceuticals Inc. Use of Cysteamine and Derivatives Thereof to Treat Mitochondrial Diseases

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"Purinergic signalling", WIKIPEDIA, 28 May 2016 (2016-05-28), XP055452858, Retrieved from the Internet <URL:https://en.wikipedia.org/wiki/Purinergic_signalling> [retrieved on 20170907] *
HAMIDPOUR ET AL.: "Antipurinergic Therapy with Suramin as a Treatment for Autism Spectrum Disorder", JOURNAL OF BIOMEDICAL SCIENCES, vol. 5, 29 March 2016 (2016-03-29), pages 1 - 7, XP055452841 *
MYHILL ET AL.: "Chronic fatigue syndrome and mitochondrial dysfunction", INT J CLIN EXP MED, vol. 2, 15 January 2009 (2009-01-15), pages 1 - 16, XP055452843 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11026904B2 (en) 2019-01-28 2021-06-08 Mitochondria Emotion, Inc. Mitofusin activators and methods of use thereof
US11083699B2 (en) 2019-01-28 2021-08-10 Mitochondria Emotion, Inc. Trans-4-hydroxycyclohexyl phenyl amide mitofusin activators and methods of use thereof
WO2020160108A1 (fr) * 2019-01-30 2020-08-06 Arizona Board Of Regents On Behalf Of The University Of Arizona Biomarqueurs lipidiques pour le dépistage et la surveillance du cancer
JP7360807B2 (ja) 2019-04-26 2023-10-13 株式会社東レリサーチセンター 疲労状態の検出を補助する方法
WO2020247127A1 (fr) * 2019-06-07 2020-12-10 Paxmedica, Inc. Compositions et méthodes pour traiter des troubles du système nerveux central
CN114839298A (zh) * 2019-12-28 2022-08-02 中精普康(北京)医药科技有限公司 一种用于检测结直肠癌或腺瘤的生物标志物及其方法
CN114839298B (zh) * 2019-12-28 2024-06-07 中精普康(北京)医药科技有限公司 一种用于检测结直肠癌或腺瘤的生物标志物及其方法

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