Human Metabolome Human Metabolome Technologies Inc. (HMT) is a leading metabolomics service provider company established on July 2003 based on capillary electrophoresis mass spectrometry (CE-MS) technologies. Technologies The company was listed on the Mothers section of Tokyo Stock Exchange in December 2013. Our main Commissioned metabolome analysis services business is commissioned metabolomics analysis using capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS): We have a time-tested track record in a number of different fields including medical sciences, pharmaceuticals, food products, fermentation, and cosmetics. With an aim of contributing in a wide range of fields, we will now set our sights on untapped areas such as the environment, energy, and chemical industries. History Jul 2003 Founded in Suehiromachi in Tsuruoka, Yamagata Prefecture with capital of 10 million yen. Jun 2004 Concluded a joint research agreement with Ajinomoto Co., Inc. May 2009 Commenced the “HMT Research Grant for Young Leaders” Aug 2012 Launched a cancer research specialized package, “C-SCOPE.” Oct 2012 Established a sales subsidiary, “Human Metabolome Technologies America, Inc.” in Massachusetts, USA Sep 2013 Registered the patent “The biomarker for depression, the measuring method for the biomarker of depression, and the program and storage for the diagnostic method” (patent number 5372213) in Japan Dec 2013 Listed on the Mothers section of the Tokyo Stock Exchange Jan 2016 Established a biomarker business company “HMT Biomedical Co., Ltd.” in Yokohama, Kanagawa, Japan May 2017 Established a sales subsidiary, “Human Metabolome Technologies Europe B.V.” in Leiden, Netherlands Apr 2018 Launched functional lipidomics specialized package “Mediator Scan” Human Metabolome Technologies Inc. ■Headquarters and Laboratory ■Human Metabolome Technologies America, Inc. 246-2 Mizukami, Kakuganji, Tsuruoka, Yamagata 24 Denby Road, Suite217 Boston, MA 02134 997-0052, Japan Phone： +1-617-871-9940 ■Tokyo Office ■Human Metabolome Technologies Europe B.V. Stern Chuo Bldg. 5F, 2-9-6 Shinkawa, Chuo, Tokyo J.H. Oortweg 21, 2333 CH Leiden, Netherlands 104-0033, Japan Phone: +31-71-332-2040 Phone: +81-3-3551-2180 Fax: +81-3-3551-2181 ■HMT BioMedical Inc. 2-11-5 Shin-Yokohama, Kohoku-ku, Yokohama 222-0033 Phone: +81-45-534-9316 Fax: +81-45-534-9317 Dealer 201810ver.4 What is Metabolomics? Application of Metabolomics Inside the body of living organisms including humans, For instance, in sickness, dietary, or environmental changes, metabolites in the living organisms (i.e., in the there exist small molecules such as amino acids, blood, urine, tissue, etc.) also change accordingly. Observation of these changes is useful for discovery of sugars, and organic acids. Most of these substances biomarkers, metabolites showing disease-specific rise and fall: we can use them to diagnose diseases, and are “metabolites” produced through enzymatic activi- also for revealing the elucidation of action mechanisms of medications, and evaluation of phenotypic effects ties. The methodology to analyze molecular species or by genetic mutations and abnormal protein function on metabolism. In the areas of APIs, fermentation, and concentrations of many metabolites at once is either production of food materials, metabolomics is expected to complement insufficiency of data pertaining to called “metabolome analysis” or “metabolomics.” product quality which gene- or protein-based assessment do not go far enough to provide. This is because Metabolites interact with each other through “metabo- metabolomics visualizes comprehensive status of low-molecular-weight compounds in samples. lism,” chemical reaction networks in living organisms. As seen in the medical field, a variety of fields including the agricultural, dietetics, environmental sciences, Metabolites produced by certain substances also as well as the chemical industries, have also found metabolomics to be a very useful tool. Examples of uses serve as precursors for subsequent reactions. are: screening of substances in agricultural and Comprehensive profiling of metabolites helps us moni- livestock products, optimization of growth condi- tor dynamics of the metabolism, and provides us with tions, evaluations in selective breeding, investigation an even better understanding of the life phenomenon. of correlations between savor and components and discovery of novel functional compounds. The range of fields to which metabolomics can be applied continues to grow. HMT is well positioned to utilize its wealth of experience in metabolome analysis in What is CE-MS? order to contribute to breakthroughs in R&D in a variety of fields. There are thought to be at least 3,000 species of metabolites between 50 to 1,000 Daltons of Metabolome Analysis molecular mass, so individual analysis of each metabolite is too time-consuming to cover the Medicine entire metabolism. Therefore, it is necessary to employ methods to measure a number of com- pounds precisely and accurately with high sensitivity at once. It is also important to select an Pathological mechanism elucidation appropriate method based on intrinsic chemical prop- Discovery of erties of metabolites such as solubility and ionicity, etc. CE-MS Water-soluble ionic substances Material etiologic factors Agricultural Production Cosmetics science Capillary Electrophoresis (CE) is a suitable method for separation of water-soluble metabolites that exist in Nucleic acids Sugar phosphates Phosphate substances Optimization of Safety assessment Selective breeding production systems Improvement of productivity Carbohydrate derived Composition effects large numbers in living organisms. The tandem connec- metabolites Organic acids Amino acids Functional assessment Ecological investigations mechanistic studies tion of CE and Mass Spectrometry (a combined capillary Amino acid byproducts Polyamines Chemistry Pharmaceuticals electrophoresis system and mass spectrometer, referred Biofuels Effects and efficacy to as “CE-MS”), which is capable of detecting tiny Biorefineries Toxicity and safety LC-MS Lipid-soluble neutral substances Companion diagnostic amount of substances, the HMT’s base technology, Food products Diagnostics markers (CDx) enables high sensitivity analysis of up to several hundred Food analysis Diagnostics development different metabolites. Based on this unique method, Lipids Fatty acids Acylcarnitines Functional food evaluation Therapeutic effects HMT conducts analysis of metabolites and develops Discovery of functional assessment Bile acids Polyphenols Steroids components new technologies. Employing CE-MS in conjunction with LC-MS based on liquid chromatography (LC) also covers a more diverse range of chemical properties. 02 HMT HMT 03 Focus further on metabolism Analysis Analysis Cost performance model targets using labeling analysis targets CE-TOFMS CE-TOFMS Roughly Quantitation 30+24 Quantitation (with units e.g. μM) (with units e.g. μM) 900 substances substances Basic Scan Relative quantitation Relative quantitation (Without units) (Without units) (Peak area value) (Peak area value) Ideal for monitoring basic metabolic changes, and analyzing mechanisms. Stable isotope labeling analysis shows the process in which labeled compounds are metabolized. Makes it possible to obtain metabolic information that serves as Ideal for deeper analysis regarding the central energy metabolism and the base for advancing research a step further. amino acid metabolism. Specifications...Time for delivery: 60 days / Analysis device: CE-TOFMS / Reports: Relative values of detected substances, Specifications...Time for delivery: 60 days / Analysis device: CE-TOFMS / Reports: Relative values of each isotopomer of group comparisons using detection data, principal component analysis, heat map, pathway map / Analysis targets: Roughly detected substances, group comparisons using detection data, pathway map / Analysis targets: 30 substances (Anion: 900 substances appearing on CE-MS annotation list (p. 10-12) / Targets (examples): Amino acids, nucleotides, nucleosides, glycolysis, tricarboxylic acid (TCA) cycle, pentose phosphate pathway, ATP, ADP, AMP, etc.), 24 substances (Cation: Amino nucleobase, Coenzyme A (CoA) derivatives, organic acids, nicotinamide coenzymes, etc. acid, citrulline, ornithine, glutathione) Optional services...Q-OPTION 110, Q-OPTION 400, Peptide Search * Labeled compounds to be added or administered must be prepared by customers. Analysis targets Example of report LC-TOFMS Roughly Analysis 〈Information on detected compounds〉 Integration of advanced technologies targets 300 substances CE-TOFMS HMT DB † Relative Area Comparative Analysis ID 0h 6h 12h 18h 6h vs 0h 12h vs 0h 18h vs 0h Roughly Compound name ¶ || ¶ || ¶ || Quantitation Mean S.D. Mean S.D. Mean S.D. Mean S.D. Ratio p-value Ratio p-value Ratio p-value 900 (with units e.g. μM) A_0003 Pyruvic acid 3.6E-04 6.8E-05 2.8E-04 5.7E-05 2.3E-04 4.6E-05 2.6E-04 N.A. 0.8 0.109 0.7 0.073 0.7 N.A. substances A_0004 Butyric acid 7.7E-05 6.2E-05 4.7E-05 9.7E-06 9.6E-05 4.4E-05 1.1E-04 5.1E-05 0.6 0.343 1.2 0.600 1.4 0.389 Dual Scan Relative quantitation (Without units) A_0005 Lactic acid 1.8E-01 3.1E-02 1.5E-01 1.1E-02 8.3E-02 2.6E-02 9.4E-02 1.8E-02 0.8 0.091 0.5 8.4E-04 *** 0.5 0.002 ** (Peak area value) A_0006 Valeric acid 8.5E-05 N.A. N.D. N.A. 1.0E-04 1.9E-05 1.2E-04 1.8E-05 <1 N.A. 1.2 N.A. 1.4 N.A. A_0007 3-Hydroxybutyric acid 3.0E-03 1.4E-03 4.0E-03 1.4E-03 1.4E-02 3.1E-03 2.4E-02 5.7E-03 1.3 0.294 4.5 6.7E-04 *** 7.8 8.9E-04 *** A_0008 2-Hydroxybutyric acid 6.5E-04 1.8E-04 4.2E-04 1.4E-04 1.1E-03 2.1E-04 1.7E-03 5.6E-04 0.6 0.061 1.7 0.006 ** 2.7 0.009 ** Adds LC-MS analysis to Basic Scan, ideal for energy metabolism screening, A_0009 2-Hydroxyisobutyric acid 1.2E-04 2.1E-05 1.2E-04 2.1E-05 9.3E-05 3.7E-05 1.2E-04 3.0E-05 1.1 0.678 0.8 0.313 1.0 0.837 A_0010 Glyceric acid 6.2E-05 1.1E-05 5.5E-05 6.6E-06 4.2E-05 2.6E-06 3.9E-05 6.0E-06 0.9 0.251 0.7 0.011 * 0.6 0.005 ** including lipid metabolism.