Biomarker detection

Abstract

To provide a method for diagnosing a subject suffering from cancer, or a predisposition thereto.SOLUTION: The method comprises detecting, in a bodily sample from a test subject, the concentration of a signature compound resulting from the metabolism of at least one sugar, and / or at least one amino acid or precursor thereof, and / or at least one polyol, present in a composition previously administered to the subject. The sugar is present in the composition at a concentration of more than 20,000 mg / 100 ml, the amino acid or precursor thereof is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration of more than 25,000 mg / 100 ml. The method further comprises comparing this concentration with a reference for the concentration of the signature compound in an individual who does not suffer from cancer. An increase or decrease in the concentration of the signature compound compared to the reference suggests that the subject is suffering from cancer or has a predisposition thereto, or provides a negative prognosis of the subject's condition.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to the detection of biomarkers, in particular to diagnose various conditions, including but not limited to cancer. The present invention relates to methods, compositions and kits for the detection of biological markers for the detection of inflammatory bowel disorders. The present invention provides diagnostic and prognostic methods for detecting cancer, such as esophageal and gastric cancer or metastatic cancer. Regarding the detection of compounds as markers. [Background technology]

[0002] Esophageal adenocarcinoma is one of the five most common cancers and is more prevalent than any other cancer in Western populations. The UK has the highest incidence of esophageal adenocarcinoma in the world. Stomach cancer is the third leading cause of cancer death in the world. The 5-year survival rates for gastric and esophageal cancer remain extremely low (13% and 18%, respectively). It is one of the worst in Europe. The key to improving cancer survival rates is earlier diagnosis. However, symptoms are nonspecific and generally shared with benign conditions. By the time of diagnosis, the disease is often at an advanced stage and has a poor prognosis. Unnecessary investigations of patients with specific symptoms result in high costs. To effectively triage patients who should undergo CT and other diagnostic modalities, There is a pressing need for non-invasive testing for patients with chronic gastrointestinal symptoms. .

[0003] Previous studies have shown an association between esophageal and gastric cancer and volatile organic compounds (VOCs). The diagnostic technique is a breath test. The researchers used gas chromatography-mass spectrometry to (GC-MS) was used to suggest the presence of an exhaled volatile organic compound (VOC) profile specific to certain cancers [4]. GC-MS is a good technique for VOC identification but is essentially semi-quantitative unless a robust calibration curve is utilized, which limits the ability to reproduce research findings across different study groups. Additionally, as it requires a significant amount of analysis time per sample, it is not, of course, a high-throughput analysis itself. Direct injection mass spectrometry such as selected ion flow tube mass spectrometry (SIFT-MS) and proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS) have the advantage of being quantitative and enabling real-time analysis [5, 6]. There is a need for a reliable non-invasive diagnostic test to identify patients suffering from cancers such as esophageal and gastric cancers. Diagnostic methods for identifying these patients with cancer are very beneficial to the patients and increase the possibility of early treatment and improved prognosis.

Summary of the Invention

Problems to be Solved by the Invention

[0004] The inventors have previously developed non-invasive tests for cancer based on the detection of trace compounds such as volatile organic compounds (VOCs) in exhaled breath. The inventors have now developed new methods and compositions that transiently induce or "stimulate" cancer by administering an optimized concentration of an oral stimulant food (e.g., beverage, capsule, or solid food), thereby generating a large amount of characteristic trace compounds (e.g., VOCs), which results in improved test performance and diagnostic and / or prognostic accuracy, leading to improved and accelerated test accuracy.

Summary of the Invention

Problems to be Solved by the Invention

[0005] Based on the detection of trace compounds such as volatile organic compounds (VOCs) in exhaled breath, the inventors have previously developed non-invasive tests for cancer. The inventors have administered an optimized concentration of an oral stimulant food (e.g., beverage, capsule, or solid food), transiently inducing or "stimulating" cancer to generate a large amount of characteristic trace compounds (e.g., VOCs), thereby achieving improved test performance and diagnostic and / or prognostic accuracy, and developing new methods and compositions that bring about improved and accelerated test accuracy. By inducing or "stimulating" cancer transiently to generate a large amount of characteristic trace compounds (e.g., VOCs), the test performance, diagnostic, and / or prognostic accuracy are improved, resulting in improved and accelerated test accuracy, which is achieved by administering an optimized concentration of an oral stimulant Thus, patients with non-specific symptoms but a high risk of esophageal and gastric cancer can be identified earlier and referred for further investigation and treatment.

Means for Solving the Problem

[0006] Therefore, in a first aspect of the present invention, there is provided a method for diagnosing a subject having cancer or its predisposition or for providing a prognosis of the subject's condition, (i) Detecting the concentration of a trace compound generated by the metabolism of at least one sugar and / or at least one amino acid or its precursor and / or at least one polyol present in a composition previously administered to a subject in a biological sample from the test subject, wherein the sugar is present in the composition at a concentration higher than 20,000 mg / 100 ml, the amino acid or its precursor is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration higher than 25,000 mg / 100 ml, and the step of detecting, and and (ii) Comparing this concentration with a reference concentration of the trace compound in an individual not suffering from cancer step Including, an increase or decrease in the concentration of the trace compound compared to the reference indicates that the subject has cancer or its predisposition, or provides a negative prognosis of the subject's condition A method is provided.

[0007] The detection step (i) detects the trace compound within 30 minutes, within 25 minutes, within 20 minutes, within 15 minutes, within 10 minutes, or within 5 minutes from the administration of a composition containing at least one sugar and / or amino acid or its precursor and / or at least one polyol. ​​​​​may include. Detection step (i) involves detecting a trace compound within 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, or 5 minutes or less from the administration of a composition containing at least one sugar and / or amino acid or its precursor and / or at least one polyol. Preferably, the detection step is performed when the composition previously administered to the test subject contains at least one sugar.

[0008] Detection step (i) involves detecting a trace compound within 30 to 60 minutes, more preferably within 30 to 55 minutes, or 30 to 50 minutes, or 30 to 45 minutes, or 30 to 40 minutes, or 35 to 60 minutes, or 35 to 55 minutes, or 35 to 50 minutes, or 3 5 to 45 minutes, or 35 to 40 minutes from the administration of a composition containing at least one sugar and / or amino acid or its precursor and / or at least one polyol. Preferably, detection step (i) further involves detecting a second trace compound within 35 to 45 minutes from the administration of a composition containing at least one sugar and / or amino acid or its precursor and / or at least one polyol. Preferably, such a detection step is performed when the composition contains at least one amino acid and / or at least one polyol. Accordingly, preferably, detection step (i) is a) detecting a trace compound within 30 to 60 minutes, more preferably within 30 to 55

[0009] minutes, or 30 to 50 minutes, or 30 to 45 minutes, or 30 to 40 minutes, a) a trace compound, at least one sugar and / or amino acid or its precursor and / or​​​ from 30 minutes up to, 25 minutes up to, 20 minutes up to, 15 minutes up to, 10 minutes up to, or 5 minutes up to, within 30 minutes, within 25 minutes, within 20 minutes, within 15 minutes, within 10 minutes, or within 5 minutes, and b) detecting the trace compound between 30 and 60 minutes, more preferably between 30 and 55 minutes, or between 30 and 50 minutes, or between 30 and 45 minutes, or between 30 and 40 minutes, or between 35 and 60 minutes, more preferably between 35 and 55 minutes, or between 35 and 50 minutes, or between 35 and 45 minutes, or between 35 and 40 minutes after administration of a composition comprising at least one sugar and / or amino acid or a precursor thereof and / or at least one polyol, and including detecting between 30 and 60 minutes, more preferably between 30 and 55 minutes, or between 30 and 50 minutes, or between 30 and 45 minutes, or between 30 and 40 minutes, or between 35 and 60 minutes,

[0010] Preferably, an increase in the concentration of the trace compound compared to a reference indicates that the subject has or has a predisposition to cancer, or provides a negative prognosis for the subject's condition. Preferably, the increase in the concentration of the trace compound is at least 10%, 20%, 30%, 40%, 50%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900% or 1000% increase in the concentration of the trace compound compared to the reference. Preferably, the sugar is at least 20,000 mg / 100 ml, at least 20,50

[0011] 0 mg / 100 ml, at least 21,000 mg / 100 ml, at least 25,00 0 mg / 100 ml, at least 50,000 mg / 100 ml or at least 75, 0 mg / 100 ml It is present at a concentration of 000 mg / 100 ml. Preferably, the sugar is about 25,000 mg / 1 00 ml concentration. Preferably, the sugar is higher than 20,000 mg / 100 ml , higher than 20,500 mg / 100 ml, higher than 21,000 mg / 100 ml, 2 5,000 mg / 100 ml, higher than 50,000 mg / 100 ml, or It is present at a concentration higher than 75,000 mg / 100 ml.

[0012] Preferably, the composition contains sugar, preferably in the range of about 20,000 mg / 100 mL to 10,0 00 mg / 100 mL, more preferably in the range of about 25,000 mg / 100 mL to 75,0 00 mg / 100 mL.

[0013] One of ordinary skill in the art will understand that the term sugar may refer to monosaccharides, disaccharides, trisaccharides, oligosaccharides, and polysaccharides or sugar alcohols. Sugars include D-glucose, D-suc rose, D-lactose, D-fructose, D-mannose, D-gluose, D-gal actose, D-xylose, D-arabinose, D-lyxose, D-ribose, D-a rose, D-altrose, D-talose, D-idose, L-arabinose, L-rham nose, L-xylulose, disaccharides, trisaccharides, oligosaccharides, and polysaccharides, sorbitol, monosaccharides with more than c4, c7, and c8, sorbitol, mannitol, maltitol, la ctitol, erythritol, and may be selected from the group consisting of.

[0014] Preferably, the sugar is glucose, sorbitol, mannose, or lactose. More preferably, the sugar is glucose, mannose, or lactose. Most preferably, The sugar is glucose or lactose.

[0015] Thus, preferably, the composition contains glucose, and preferably, the glucose is present in the composition at a concentration of at least 25,000 mg / 100 ml. More preferably, the sugar is glucose and is present in the composition at a concentration of at least 25,000 mg / 100 ml, and the trace compound is detected within 10 minutes from the administration of the composition containing glucose.

[0016] The composition to be administered to the subject may contain citric acid. This may be instead of or in addition to the sugar. Preferably, citric acid is used in combination with the sugar. Preferably, the sugar is glucose. Thus, preferably, the composition contains citric acid and glucose.

[0017] Preferably, citric acid is present in the composition at a concentration of at least 1,000 mg / 100 ml, at least 1,1 00 mg / 100 ml, at least 1,200 mg / 100 ml, at least 1,300 mg / 100 ml, or at least 1,400 mg / 100 ml. Preferably, citric acid is present at a concentration of about 1,400 mg / 100 ml.

[0018] Thus, preferably, the composition contains glucose and citric acid, and preferably, the glucose is present in the composition at a concentration of at least 25,000 mg / 100 ml and the citric acid is present in the composition at a concentration of at least 1,400 mg / 100 ml. More preferably, the composition contains glucose present in the composition at a concentration of at least 25,000 mg / 100 ml and citric acid present in the composition at a concentration of at least 1,400 mg / ml. ​​​​and the trace compounds are determined to be present within 10 minutes of administration of the composition comprising glucose and citric acid. It is detected.

[0019] In another embodiment, the composition preferably contains at least 500 mg / 100 ml, at least At least 1000mg / 100ml, at least 2000mg / 100ml, at least 30 00mg / 100ml, at least 4000mg / 100ml, at least 5000mg / 100ml or contain amino acids at a concentration of at least 6000mg / 100ml is preferred.

[0020] Preferably, the composition has a concentration of 1000 mg / 100 ml, preferably greater than 500 mg / 100 ml. Higher than 100ml, higher than 2000mg / 100ml, higher than 3000mg / 100ml higher than 4000mg / 100ml, higher than 5000mg / 100ml, and contains amino acids at concentrations greater than 6000mg / 100ml.

[0021] Preferably, the amino acid is 500 mg / 100 ml to 10,000 mg / 100 ml. Between 500mg / 100ml and 6000mg / 100ml, 500mg / 100ml Between ~5000mg / 100ml, 500mg / 100ml~4000mg / 100ml Between 500mg / 100ml and 3000mg / 100ml, Between 500mg / 100ml and 3000mg / 100ml Between 1-2500mg / 100ml, 500mg / 100ml-2000mg / 100ml l, between 1000mg / 100ml and 10000mg / 100ml, between 1500mg / Between 100ml-10000mg / 100ml, 2000mg / 100ml-10000 mg / 100ml, between 2500mg / 100ml and 10000mg / 100ml, Between 3000 mg / 100 ml and 10000 mg / 100 ml, 4000 mg / 100 m l and 10000 mg / 100 ml, between 5000 mg / 100 ml and 10000 mg / 1 00 ml, between 6000 mg / 100 ml and 10000 mg / 100 ml, 1000 mg / 100 ml and 5000 mg / 100 ml, between 1000 mg / 100 ml and 300 0 mg / 100 ml, between 1000 mg / 100 ml and 2500 mg / 100 ml, between 1000 mg / 100 ml and 2000 mg / 100 ml, 1500 mg / 100 ml and 10000 mg / 100 ml, between 1500 mg / 100 ml and 5000 mg / 100 ml, between 1500 mg / 100 ml and 3000 mg / 100 ml, 1500 mg / 100 ml and 2500 mg / 100 ml, or between 1500 mg / 100 ml and 200 0 mg / 100 ml, and is present in the composition at a concentration within this range.

[0022] Preferably, the amino acid is present in the composition at a concentration of about 2000 mg / ml.

[0023] The amino acid may be selected from the group consisting of tyrosine, glutamic acid, glutamate, phenylalanine, trypt ophan, proline, and histidine.

[0024] Preferably, when the amino acid is glutamic acid, the concentration of the amino acid is at least 5,0 00 mg / 100 ml, at least 5,100 mg / 100 ml, at least 5,200 mg / 100 ml, at least 5,300 mg / 100 ml, at least 5,400 mg / 100 ml, at least 5,500 mg / 100 ml , at least 6000 mg / 100 ml, greater than 5,000 mg / 100 ml High, higher than 5,100 mg / 100 ml, higher than 5,200 mg / 100 ml, 5 ,300 mg / 100 ml, higher than 5,400 mg / 100 ml, 5,500 mg / 1 00 ml, higher than, or higher than 6,000 mg / 100 ml. Preferably, when the amino acid is glutamic acid, the concentration of the amino acid is 1,800 mg / 100 ml to 2,200 mg / 100 ml, between 1,900 mg / 100 ml and 2,100 mg / 100 ml is. Preferably, when the amino acid is glutamic acid, the concentration of the amino acid is 1,900 mg / 100 ml, 2,000 mg / 100 ml, 2,100 mg / 100 ml, 2,2 00 mg / 100 ml or 2,300 mg / 100 ml. Preferably, the amino acid is glutamic acid, the concentration of the amino acid is 2,100 mg / ml. However, in one embodiment, the amino acid is not glutamic acid.

[0025] Most preferably, the amino acid is tyrosine.

[0026] Therefore, preferably, the composition contains tyrosine, and preferably, tyrosine is at least also present in the composition at a concentration of 2,000 mg / 100 ml. More preferably, the amino acid is tyrosine and is present in the composition at a concentration of at least 2,000 mg / 100 ml, and the trace compound is detected between 35 and 45 minutes after administration of the composition containing tyrosine.

[0027] The composition administered to the subject may contain an amino acid precursor. This may be instead of, and / or in addition to, the amino acid and / or sugar. Preferably, the amino acid precursor is phenylalanine. Preferably, the amino acid precursor is in a ratio with its respective amino acid It is used in the combination of. Therefore, preferably, the composition contains tyrosine and phenylalanine. It contains.

[0028] Preferably, the amino acid precursor is at least 500 mg / 100 ml, at least 1000 mg / 100 ml, at least 2000 mg / 100 ml, at least 3000 mg / 100 ml, at least 4000 mg / 100 ml, or at least 5000 mg / 100 ml present in the composition. Preferably, the amino acid precursor is at least 500 mg / 100 ml, at least 1000 mg / 100 ml, at least 2000 mg / 100 ml, at least 3000 mg / 100 ml, at least 4000 mg / 100 ml, or at least 5000 mg / 100 ml present in the composition. Preferably, the amino acid precursor is between 500 mg / 100 ml and 10000 mg / 100 ml, between 500 mg / 100 ml and 5000 mg / 100 ml, between 500 mg / 100 ml and 4000 mg / 100 ml, between 500 mg / 100 ml and 3000 mg / 100 ml, between 500 mg / 100 ml and 2500 mg / 100 ml, between 500 mg / 100 ml and 2000 mg / 100 ml, between 1000 mg / 100 ml and 10000 mg / 100 ml, between 1500 mg / 100 ml and 10000 mg / 100 ml, between 2000 mg / 100 ml and 10000 mg / 100 ml, between 2500 mg / 100 ml and 10000 mg / 100 ml, between 3000 mg / 100 ml and 10000 mg / 100 ml, between 1000 mg / 100 ml and 5000 mg / 100 ml, between 1000 mg / 100 ml and 3000 mg / 100 ml, between 1000 mg / 100 ml and 2500 mg / 100 ml. / 100 ml, at least 2000 mg / 100 ml, at least 3000 mg / 100 ml, at least 4000 mg / 100 ml, or at least 5000 mg / 100 ml of the concentration is present in the composition. Preferably, the amino acid precursor is at least 5 00 mg / 100 ml, at least 1000 mg / 100 ml, at least 2000 mg / 100 ml, at least 3000 mg / 100 ml, at least 4000 mg / 100 ml, or at least 5000 mg / 100 ml of the concentration is present in the composition. Preferably Preferably, the amino acid precursor is between 500 mg / 100 ml and 10000 mg / 100 ml, between 500 mg / 100 ml and 5000 mg / 100 ml, between 500 mg / 100 ml and 4 000 mg / 100 ml, between 500 mg / 100 ml and 3000 mg / 100 ml between, between 500 mg / 100 ml and 2500 mg / 100 ml, between 500 mg / 100 ml and 2000 mg / 100 ml, between 1000 mg / 100 ml and 10000 mg / 100 m l, between 1500 mg / 100 ml and 10000 mg / 100 ml, between 2000 mg / 100 ml and 10000 mg / 100 ml, between 2500 mg / 100 ml and 10000 mg / 100 ml, between 3000 mg / 100 ml and 10000 mg / 100 ml, between 1000 mg / 100 ml and 5000 mg / 100 ml, between 1000 mg / 100 ml and 3000 mg / 100 ml, between 1000 mg / 100 ml and 2500 mg / 100 m between 1000 mg / 100 ml and 2000 mg / 100 ml, between 1500 mg / 1 00 ml and 10000 mg / 100 ml, between 1500 mg / 100 ml and 5000 mg / 100 ml, between 1500 mg / 100 ml and 3000 mg / 100 ml, between 150 0 mg / 100 ml and 2500 mg / 100 ml, or present in the composition at a concentration between 1500 mg / 100 ml and 2000 mg / 100 ml.

[0029] Preferably, the amino acid precursor is phenylalanine. Preferably, phenylalanine is present at a concentration of 3000 mg / 100 ml.

[0030] Preferably, the composition contains phenylalanine and tyrosine.

[0031] In one embodiment, the composition contains tyrosine, phenylalanine and glutamic acid. Preferably, tyrosine is present at a concentration of at least 2,000 mg / 100 ml, and phenyl alanine is present at a concentration of at least 3,000 mg / 100 ml, and glutamic acid is at least present at a concentration of 2,100 mg / 100 ml.

[0032] Preferably, the polyol is present in the composition at a concentration higher than 25,000 mg / 100 ml. Preferably, the polyol is at a concentration higher than 26,000 mg / 100 ml, higher than 27,0 00 mg / 100 ml, higher than 28,000 mg / 100 ml, or higher than 29, 000 mg / 100 ml in the composition. Preferably, the polyol is , higher than 30,000 mg / 100 ml, higher than 35,000 mg / 100 ml, 4 0,000 mg / ml, higher than 45,000 mg / 100 ml, 50,000 It is present in the composition at a concentration higher than mg / 100ml. Preferably, the polyol is less than or equal to 30,000 mg / 100 ml, at least 35,000 mg / 100 ml, at least 40,000 mg / ml, at least 45,000 mg / 100 ml, at least 50,000 mg / 100 ml and is present in the composition.

[0033] Preferably, the polyol is present in the composition at a concentration of 50,000 mg / 100 ml. Most preferably, the polyol is between 23,000 mg / 100 ml and 27,000 mg / 100 ml, or between 24,000 mg / 100 ml and 26,000 mg / 100 m l and is present in the composition.

[0034] Preferably, the polyol is glycerol. Preferably, glycerol is at a concentration higher than 30, 000 mg / ml, more preferably 50,000 mg / 100 ml and is present in the composition. Most preferably, glycerol is between 23,000 mg / 100 ml and 2 7,000 mg / 100 ml, or between 24,000 mg / 100 ml and 26,000 mg / 100 ml and is present in the composition.

[0035] In one embodiment, at least one sugar and / or at least one amino acid or its precursor and / or at least one polyol is metabolized by cancer-related microorganisms.

[0036] "Prognosis" may be relevant to the determination of the treatment outcome in a subject diagnosed with cancer. The prognosis is related to the rate and / or duration of cancer progression or improvement in the subject. ​、may be related to predicting survival probability and / or the effectiveness of various treatment regimens Thus, poor prognosis may indicate cancer progression, low survival probability, and reduced effectiveness of treatment regimens. Good prognosis may indicate cancer improvement, high survival probability, and increased effectiveness of treatment regimens.

[0037] Cancer-related microorganisms may be bacteria. It is understood that the microorganisms and bacteria present in the intestine form the so-called "microbiome". Thus, cancer-related microorganisms that metabolize at least one substrate into trace compounds detected and / or analyzed by the method of the present invention for diagnosing cancer preferably form part of the microbiome.

[0038] Cancer-related microorganisms may be Streptococcus, Lactobacillus, Ve illonella, Prevotella, Neisseria, Haemophil us, L.coleohominis, Lachnospiraceae, Klebsi ella, Clostridiales, Erysipelotrichales or any combination thereof.

[0039] Cancer-related microorganisms may be S.pyogenes, Klebsiella pneumoni ae, Lactobacillus acidophilus or any combination thereof.

[0040] Cancer-related microorganisms may be E. coli, P. mirabili, B. cepacia 、S. pyogenes, Streptococcus salivarius, Ac ​​​​Tinomyces naeslundii, Lactobacillus ferme ntum, Streptococcus anginosus, Clostridium bifermentans, Clostridium perfringens, Cl ostridium septicum, Clostridium sporogene s, Clostridium tertium, Eubacterium lentum , Eubacterium sp., Fusobacterium simiae, Fu sobacterium necrophorum, Lactobacillus ac idophilus, Peptococcus niger, Peptostrepto coccus anaerobius, Peptostreptococcus asa ccharolyticus, Peptostreptococcus prevoti i, P. aeruginosa, S. aureus, P. mirabilis, E . faecalis, S. pneumoniae, N. meningitides , Acinetobacter baumannii, Bacteroides cap illosus, Bacteroides fragilis, Bacteroides pyogenes, Clostridium difficile, Clostrid ium ramosum, Enterobacter cloacae, Klebsie lla pneumoniae, Nocardia sp., Propionibact erium acnes, Propionibacterium propionicu m or any combination thereof. Preferably, the cancer-related microorganism is E. coli, L. fermentum, S. salivarius, S. angin osus or K. pneumoniae.

[0041] In an embodiment, the cancer is gastroesophageal junction cancer, gastric cancer, esophageal cancer, esophageal squamous cell carcinoma (E SCC), or esophageal adenocarcinoma (EAC). Thus, in a preferred embodiment, the diagnosis is for diagnosing gastroesophageal junction cancer, gastric cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), or esoph ageal adenocarcinoma (EAC). Most preferably, the cancer is a gastroesophageal cancer such that this condition can be diagnosed or prognosed. The cancer may be metastatic.

[0042] Preferably, the cancer is gastric cancer, esophageal cancer or metastatic cancer.

[0043] In an embodiment, the cancer is pancreatic cancer or colorectal cancer. Thus, the diagnosis or prognosis may also be for diagnosing or prognosing pancreatic cancer or colorectal cancer.

[0044] In a second aspect, a method for detecting a trace compound in a test subject, comprising: (i) providing to the subject a composition comprising at least one substrate according to the first aspect that becomes a trace compound, and and (ii) detecting the concentration of the trace compound in a bodily sample from the subject is provided.

[0045] Preferably, the detecting step is performed according to the first aspect.

[0046] In a third aspect of the invention, preferably for use in a method for diagnosing or prognosticating cancer, ​​A composition comprising at least one sugar and / or at least one amino acid or its precursor and / or at least one polyol, which is suitable for metabolism into trace compounds, wherein the sugar is present in the composition at a concentration higher than 20,000 mg / 100 ml, the amino acid is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration higher than 25,000 mg / 100 ml.

[0047] Preferably, the composition and the cancer are as defined in the first aspect.

[0048] In a fourth aspect, there is provided a composition comprising at least one substrate suitable for metabolism into trace compounds by cancer-related microorganisms for use in the method of the first or second aspect.

[0049] In a fifth aspect, there is provided a kit for diagnosing a subject having cancer or its predisposition, or for providing a prognosis of the subject's condition, the kit comprising: (a) a composition comprising at least one substrate as defined in the first aspect, (b) means for determining the concentration of trace compounds in a sample from the test subject, and (c) a reference of the concentration of trace compounds in a sample from an individual not having cancer, wherein the kit is used to identify an increase or decrease in the concentration of trace compounds in a biological sample from the test subject compared to the reference, thereby indicating that the subject has cancer or its predisposition, or providing a negative prognosis of the subject's condition.

[0050] Preferably, the composition and the cancer are as defined in the first aspect.

[0051] ​​​​​​​​​The methods of the first and second aspects prevent, reduce, or delay cancer progression and include administering or causing to be administered a therapeutic agent to a subject, or causing the subject to follow a special diet, or performing chemotherapy or chemoradiation therapy.

[0052] Accordingly, in a sixth aspect, a method of treating a subject having cancer, (i) providing to the subject a composition comprising at least one substrate defined in the first aspect, (ii) analyzing the concentration of a trace compound produced by the metabolism of at least one substrate in a bodily sample from the test subject and comparing this concentration to a reference concentration of the trace compound in an individual not having cancer, wherein an increase or decrease in the concentration of the trace compound in the bodily sample from the test subject compared to the reference indicates that the subject has cancer or a predisposition thereto, or has a negative prognosis, and (iii) administering or causing to be administered a therapeutic agent to the subject, or causing the subject to follow a special diet, or performing chemotherapy or chemoradiation therapy, wherein the therapeutic agent or special diet, or chemotherapy or chemoradiation therapy prevents, reduces, or delays cancer progression, is provided. Preferably, the composition and the cancer are as defined in the first aspect.

[0053] The methods of the invention are useful for monitoring the effectiveness of treating related cancers.

[0054] For example, the treatment of resectable esophagogastric cancer is neoadjuvant chemotherapy, or chemoradiation It may include surgery and adjuvant chemotherapy following the therapy. For very early esophageal gastric cancer treatment may include endoscopic resection. Treatment for advanced esophageal gastric cancer may include palliative chemotherapy (Recently, it has been shown that the cancer-related microbiome enhances liver metastasis (Bullman et al., Science, 2017). Therefore, using the present invention described herein it is possible to monitor the response to cancer-related microbiome-directed therapy.

[0055] If the cancer is pancreatic cancer, treatment may include administering chemotherapy, chemoradiotherapy with or without surgery. For example, if the cancer is colorectal cancer, treatment may include administering chemotherapy, chemoradiotherapy with or without surgery, or endoscopic resection.

[0056] In a seventh aspect, a method for determining the effectiveness of treating a subject suffering from cancer by a therapeutic agent or a special diet, or by chemotherapy or chemoradiotherapy, comprising: (i) providing to the subject a composition comprising at least one substrate according to the first aspect; and (ii) analyzing the concentration of a trace compound produced by the metabolism of at least one substrate in a body sample from the test subject and comparing this concentration to a reference concentration of the trace compound in an individual not suffering from cancer; wherein an increase or decrease in the concentration of the trace compound in the body sample from the test subject compared to the reference indicates whether the treatment regimen by the therapeutic agent or special diet, or by chemotherapy or chemoradiotherapy, is effective or ineffective. A method is provided.

[0057] Preferably, the composition and the cancer are as defined in the first aspect.

[0058] The composition may be an existing composition, food or beverage containing any one of the aforementioned components. Preferably, the composition contains water. The composition of the present invention is ingested by a subject. The composition may be a solid or fluid that can be eaten or swallowed. In an embodiment, the composition can be chewed, thereby causing the release of the substrate, which descends to the intestine. In an implementation form, the composition may be in the form of a capsule designed to decompose at a specific location by the digestive tract, thereby resulting in the targeted release of at least one substrate. However, the composition is preferably a liquid (i.e., a beverage) that can be swallowed, which may be referred to as an oral stimulating drink (OSD).

[0059] Preferably, a sample is taken from the subject, and then trace compounds in the body sample are detected. In some embodiments, the concentration of the trace compound is measured.

[0060] The trace compound may be any compound that may indicate or correlate with the presence of microorganisms. The detected trace compounds may be volatile organic compounds ( VOCs) that result in a fermentation profile, and these may be detected in body samples by various techniques. In one embodiment, these compounds may be detected in a liquid or semi-solid sample in which they are dissolved. However, in a preferred embodiment, the compounds are detected from a gas or vapor. For example, if the trace compounds are VOCs, they may be emitted from the sample or form part of it, and thus can be detected in the form of a gas or vapor.

[0061] An increase or decrease in the concentration of these trace compounds compared to a reference indicates that the subject has cancer or has a predisposition thereto, or provides a negative prognosis for the subject's condition. Preferably, an increase in the concentration of these trace compounds compared to a reference indicates that the subject has cancer or has a predisposition thereto, or provides a negative prognosis for the subject's condition. or has a predisposition thereto, or provides a negative prognosis for the subject's condition. An increase in the concentration of these trace compounds compared to a reference indicates that the subject has cancer or has a predisposition thereto, or provides a negative prognosis for the subject's condition. or has a predisposition thereto, or provides a negative prognosis for the subject's condition. or provides a negative prognosis for the subject's condition.

[0062] The VOCs may be short-chain fatty acids, aldehydes, alcohols, or any combination thereof. or any combination thereof.

[0063] The VOCs may be C 1 ~C 3 aldehydes, C 1 ~C 3 alcohols, C ~C 2 ~C 10 alkanes, C 1 ~C 20 alkanes, C 4 ~C 10 alcohols, C 1 ~C 6 carboxylic acids, C 4 ~C 20 aldehydes, optionally C 1 ~C 6 phenols substituted with an alkyl group, C 2 aldehydes, C 3 alde hyde, C 8 aldehydes, C 9 aldehydes, C 10 aldehydes, C 11 aldehydes, any analogues or derivatives of the aforementioned species, or any combination thereof.

[0064] C 1 ~C 6The carboxylic acid may be selected from the group consisting of formic acid, acetic acid, propanoic acid, butanoic acid, pentanoic acid, and hexanoic acid. C 1 ~C 3 The aldehyde may be selected from the group consisting of formaldehyde, acetaldehyde, and propanal. C 4 ~C 20 a ldehyde may be C 4 ~C 10 aldehyde. C 4 ~C 20 The aldehyde may be buta nal, pentanal, hexanal, heptanal, octanal, nonanal, deca nal, undecanal, dodecanal, tridecanal, tetradecanal, pentadeca nal, hexadecanal, heptadecanal, octadecanal, nonadecanal and icodanal. C 1 ~C 20 The alkane is preferably C ~C 4 ~C 16 alkane, more preferably C 8 ~C 14 alkane. C 1 ~C 20 The alkane may be methane, ethane, propane, butane, pentane, hexane, he ptane, octane, nonane, decane, undecane, dodecane, tridecane, tetradecane , pentadecane, hexadecane, heptadecane, octadecane, nonadecane, and ico dane. The phenol may be unsubstituted. Alternatively, the phenol may be substituted with a C alkyl group at the trans position in the range of C 1 ~C 6 The phenol may be substituted with a C 1 ~C 3 alkyl group. Optionally, C1 ~C 6 substituted with an alkyl group The phenol may be phenol, 1-hydroxy-4-ethylbenzene or p-cresol and may be.

[0065] Preferably, the volatile organic compound (VOC) is acetic acid, butanoic acid, hexanoic acid, pentane acid, propanoic acid, acetaldehyde, decanal, heptanal, hexanal, nonanal , octanal, pentanal, butanal, propanal, 1-hydroxy-4- ethylbenzene, decane, dodecane, p-cresol and phenol or any of these selected from the group consisting of combinations.

[0066] When the substrate is a sugar, preferably glucose, the trace compound is acetic acid, butanoic acid, penta nic acid, propanoic acid, hexanoic acid, acetaldehyde, propanal, butanal, hexa nal, pentanal, decanal, 1-hydroxyethylbenzene (1-hydoxytheylben zene) and / or p-cresol and may be.

[0067] When the substrate is a sugar, preferably glucose, acetic acid, butanoic acid, pentanoic acid, propano ic acid, acetaldehyde, butanal, hexanal, pentanal, 1-hydroxyethyl benzene and / or an increase in p-cresol may indicate gastric cancer. Preferably , when the substrate is glucose and the trace compound is butanoic acid, an increase in the concentration of the trace compound is an increase of at least 300% in the concentration of the butanoic acid compound compared to the reference and indicates gastric cancer . Preferably, when the substrate is glucose and the trace compound is propanoic acid, the trace An increase in the concentration of the trace compound is at least 100% of the concentration of the propanoic acid compound compared to the reference, indicating gastric cancer. Preferably, when the substrate is glucose and the trace compound is acetic acid , the increase in the concentration of the trace compound is at least 200% of the concentration of the acetic acid compound compared to the reference, indicating gastric cancer. Preferably, when the substrate is glucose and the trace compound is pentanoic acid, the increase in the concentration of the trace compound is at least 50% of the concentration of the pentanoic acid compound compared to the reference, indicating gastric cancer. When the substrate is a sugar, preferably glucose, an increase in acetic acid, pentanoic acid, propanoic acid, butanol , propanal and / or hexanoic acid may indicate esophageal cancer. Preferably, when the substrate is glucose and the trace compounds are butanoic acid, propanoic acid and / or

[0068] acetic acid, the increase in the concentration of the trace compounds is at least 50% of the concentration of the butanoic acid, propanoic acid and / or acetic acid compounds compared to the reference, indicating esophageal cancer. When the substrate is a sugar, preferably glucose, and in combination with citric acid, an increase in the trace compounds butanoic acid, propanoic acid and / or propanal may indicate esophageal cancer .

[0069] When the substrate is a sugar, preferably glucose, and in combination with citric acid, an increase in the trace compounds butanoic acid, propanoic acid and / or propanal may indicate gastric cancer. .

[0070] When the substrate is an amino acid or its precursor, the trace compounds are butanal, decanal , heptanal, hexanal, phenol, decane, p-cresol, 1-hydroxy

[0071] butane, etc. butane, etc. It may be ethylbenzene and / or dodecane. Preferably, when the substrate is an amino acid or its precursor, the increase in the concentration of the trace compound is at least 10%, 20%, 30%, 40% or 50% increase compared to the reference. When the substrate is tyrosine, the trace compound may be butanal, decanal, heptanal, hexanal, phenol, decane, p-cresol and / or dodecane.

[0072] Preferably, the trace compound is decanal and / or dodecane. Preferably, when the substrate is tyrosine, the increase in the concentration of the trace compound is at least 10%, 20%, 30%, 40% or 50% increase compared to the reference. When the substrate is tyrosine, an increase in decanal may indicate esophageal cancer.

[0073] When the substrate is tyrosine, an increase in dodecane may indicate gastric cancer.

[0074]

[0075] When the substrate is phenylalanine, the trace compound may be dodecane, decane, phenol, decanal and / or dodecane.

[0076] When the substrate is phenylalanine, an increase in the trace compounds decanal, 1-hydroxyethylbenzene, decane, dodecane, p-cresol and / or phenol may indicate esophageal cancer.

[0077] When the substrate is phenylalanine, an increase in the trace compounds hydroxyethylbenzene, decane, dodecane, p-cresol and / or phenol may indicate gastric cancer.

[0078] ​​​​​​​​​ When the substrate is glutamic acid, the trace compounds may be propanal, dodecane, phenol and / or butanoic acid.

[0079] When the substrate is glutamic acid, an increase in the trace compounds propanal, dodecane, phenol and / or butanoic acid may indicate esophageal cancer.

[0080] When the substrate is glutamic acid, an increase in the trace compounds propanal, dodecane, phenol and / or butanoic acid may indicate gastric cancer.

[0081] When the substrate is a polyol, preferably glycerol, the trace compounds may be butanoic acid, acetic acid, hexanoic acid, pentanoic acid, propanoic acid, butanal, hexanal, pentanal and / or propanal.

[0082] When the substrate is a polyol, preferably glycerol, an increase in the trace compounds butanoic acid, acetic acid, hexanoic acid, pentanoic acid, propanoic acid, butanal, hexanal, pentanal and / or propanal may indicate esophageal cancer.

[0083] When the substrate is a polyol, preferably glycerol, an increase in the trace compounds butanoic acid, acetic acid, hexanoic acid, pentanoic acid, propanoic acid, butanal, hexanal, pentanal and / or propanal may indicate gastric cancer.

[0084] Preferably, the sample is any body sample in which trace compounds are present or secreted. Thus, preferably the detection or diagnostic method is performed in vitro. However, the prognostic method may be performed in vivo. For example, the sample may include urine, feces, hair , sweat, saliva, blood or tears. In one embodiment, the sample may be immediately assayed for the level of trace compounds. Alternatively, the sample may be stored at a low temperature, e.g., in a freezer, until the concentration of the trace compounds is determined, or even frozen . The measurement of trace compounds in a body sample may be made on the whole sample or a processed sample, e.g., whole blood or processed blood .

[0085] In an embodiment, the sample may be a urine sample. The concentration of trace compounds in a body sample is preferably measured in vitro from a urine sample collected from the subject . The compound may be detected from the gas or vapor emitted from the urine sample. It is understood that the detection of compounds in the gas phase emitted from urine is preferred .

[0086] It is also understood that a "fresh" body sample may be analyzed immediately after collection from the subject . Alternatively, the sample may be frozen and stored. The sample may then be thawed and analyzed at a later date .

[0087] However, most preferably, the body sample may be an exhaled breath sample from the test subject. The sample is preferably collected from the subject's mouth after nasal inhalation. Preferably, the sample contains the alveolar air of the subject. Preferably, the alveolar air is collected beyond the dead space air by capturing the end-expired breath . Thereafter, the VOC from the exhaled breath bag is preferably preconcentrated into the tube by moving the exhaled breath across a thermal desorption tube .

[0088] The concentration difference of trace compounds indicating cancer or its predisposition in the subject is an increased compared to a reference or may be decreased. The concentration of a trace compound in a patient suffering from a disease depends highly on many factors, such as how far the disease has progressed, as well as the age and gender of the subject. It is understood. Also, the reference concentration of a trace compound in an individual not suffering from a disease may vary to some extent but, on average, over a given period, the concentration tends to be substantially constant. Furthermore, it should be understood that the concentration of a trace compound in one group of individuals suffering from a disease may be different from the concentration of the compound in another group of individuals not suffering from the disease. However, it is possible to determine the average concentration of a trace compound in an individual not suffering from cancer, which is referred to as the reference or "normal" concentration of the trace compound. The normal concentration corresponds to the reference value described above. In one embodiment, the method of the present invention preferably determines the ratio of chemicals in exhaled breath (i.e., using other components therein as a reference) and compares the markers for these diseases to indicate whether they are elevated or reduced.

[0089] In one embodiment, the method of the present invention preferably determines the ratio of chemicals in exhaled breath (i.e., using other components therein as a reference) and compares the markers for these diseases to indicate whether they are elevated or reduced. including.

[0090] Trace compounds are preferably volatile organic compounds (VOCs) that provide a profile and can be detected in or from a body sample by various techniques. Thus they may be detected using a gas analyzer. Examples of suitable detectors for detecting trace compounds include, preferably, electrochemical sensors, semiconductor metal oxide sensors, water crystal oscillator microbalance sensors, optical dye sensors, fluorescence sensors, conductive polymer sensors , composite polymer sensors, or optical spectroscopy. crystal oscillator microbalance sensors, optical dye sensors, fluorescence sensors, conductive polymer sensors , composite polymer sensors, or optical spectroscopy.

[0091] The inventors used gas chromatography, mass spectrometry, GCMS or TOF and demonstrated that trace compounds can be reliably detected. A dedicated sensor may be used in the detection step .

[0092] The reference value may be obtained by assaying a statistically significant number of control samples (i.e., samples from subjects without the disease). Thus, the kit of the fifth aspect of the present invention may be a control sample (for assay).

[0093] The device preferably includes a positive control corresponding to the trace compound (most preferably provided in a container). The device preferably includes a negative control (preferably provided in a container). In a preferred embodiment, the kit may include a reference, a positive control and a negative control. The kit may also include additional controls such as "spike-in" controls that provide concentration references as needed, and additional positive controls for each of the trace compounds, or analogs or derivatives thereof.

[0094] Thus, the inventors recognized that the concentration difference of the trace compound between the reference normal (i.e., control) level and the increased / decreased level can be used as a physiological marker indicating the presence of a disease in the test subject. If the subject has an increase / decrease in the concentration of one or more trace compounds that is significantly higher / lower than the control value that is the "normal" concentration of the compound in the reference, it is understood that the subject is at high risk of having a disease or a more advanced condition compared to the case where the concentration of the compound is only slightly higher / lower than the "normal" concentration.

[0095] ​​​​​​​​​One skilled in the art would understand a method of measuring the concentration of trace compounds in a statistically significant number of control individuals and the concentration of the compounds in the test subject, using each numerical value to determine whether the test subject has a statistically significant increase / decrease in the concentration of the compound, and thus inferring whether the subject suffers from the disease being screened. The kit of the fifth aspect may include sample extraction means for obtaining a sample from the test subject. The sample extraction means may include a needle or a syringe, etc. The kit may include a sample collection container for receiving an extraction sample that can be liquid, gaseous or semi-solid. The kit may further include instructions for use. In a further aspect, a method for diagnosing a subject suffering from cancer or its predisposition, or for providing a prognosis of the condition of the subject, comprising:

[0096] (i) Detecting the concentration of trace compounds produced by the metabolism of at least one sugar, at least one amino acid or its precursor, and / or at least one polyol present in a composition previously administered to the subject in a body sample from the test subject, wherein the sugar is present in the composition at a concentration higher than 20,000 mg / 100 ml, the amino acid or its precursor is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration higher than 30,000 mg / 100 ml, the step of detecting, and (ii) Comparing this concentration with a reference concentration of trace compounds in individuals not suffering from cancer step, and an increase or decrease in the concentration of the trace compound compared to the reference indicates that the subject has cancer

[0097] (i) Detecting the concentration of trace compounds produced by the metabolism of at least one sugar, at least one amino acid or its precursor, and / or at least one polyol present in a composition previously administered to the subject in a body sample from the test subject, wherein the sugar is present in the composition at a concentration higher than 20,000 mg / 100 ml, the amino acid or its precursor is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration higher than 30,000 mg / 100 ml, the step of detecting, and (ii) Comparing this concentration with a reference concentration of trace compounds in individuals not suffering from cancer step, and an increase or decrease in the concentration of the trace compound compared to the reference indicates that the subject has cancer (ii) Comparing this concentration with a reference concentration of trace compounds in individuals not suffering from cancer step including, and an increase or decrease in the concentration of the trace compound compared to the reference indicates that the subject has cancer ​​​​​​​or suggest having such predisposition, or provide a negative prognosis of the condition of the subject A method is provided.

[0098] In another aspect, at least one sugar and / or at least one amino acid or its precursor and / or at least one polyol present in the composition, preferably for use in a method for diagnosing or prognosticating cancer, are suitable for metabolism to a trace compound A composition is provided, wherein the sugar is present in the composition at a concentration higher than 20,000 mg / 100 ml, the amino acid is present in the composition at a concentration of at least 500 mg / ml, and the polyol is present in the composition at a concentration higher than 30,000 mg / 100 ml and / or at least one polyol, and the composition contains

[0099] All features described herein (including any appended claims, abstract, and drawings) and / or any steps of any method or process thus disclosed may be combined in any combination with any of the above aspects, except combinations where at least some of such features and / or steps are mutually exclusive

[0100] For a better understanding of the present invention and to show how embodiments of the present invention may be carried out, reference is now made, by way of example, to the accompanying drawings BRIEF DESCRIPTION OF THE DRAWINGS

[0101]

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Mode for Carrying Out the Invention

[0102] Materials and Methods [Example 1] Glucose Dosage Test Subjects Four healthy subjects volunteered to participate and gave written informed consent.

[0103] Dosage Concentration Four dosages of the substrate were derived from (i) the recommended daily intake level by the Food and Nutrition Board, and (ii ) an established glucose tolerance test. In the glucose tolerance test, 1 Use 75 g of glucose, which is acceptable, dissolved in 100 ml of water, which is good for the patient. The maximum recommended daily dose is 130 g per day for adults. [1] Based on these findings, the inventors selected doses of 75 g, 50 g, 25 g, 10 g to compare the dose response to glucose concentration. All

[0104] Exhaled breath sampling The method for detecting short-chain fatty acids was established by Selective Ion Flow Tube Mass Spectrometry (SIFT-MS VoiceUl tra 200; Syft Technologies, Anatune, UK). All exhaled breath sampling was performed in the morning, and subjects maintained a clear liquid diet for at least 6 hours prior to exhaled breath sampling. All subjects exhaled directly into the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled breath test was performed for each method, followed by consumption of glucose dissolved in 100 ml of warm water, followed by rinsing the mouth 3 times with water to decontaminate the oral cavity. Direct sampling of 3

[0105] exhaled breath samples over 60 seconds was performed continuously at 5- minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all 4 methods. into the flow tube, reacting with the precursor ions to generate product ions, Separate according to. SIFT-MS was subjected to an automated verification cycle once a day to operate within a temperature range of 10 to 30 °C. The data was obtained in parts per billion concentration units.

[0106] Sugar Comparison of four different sugars at a dose of 25 g each. 25 g was selected after the first glucose test where similar VOC concentrations were observed between 25 g and 75 g. Glucose, lactose and mannose followed a similar pattern with a maximum increase occurring 10 minutes after sugar consumption (Figure 16). Lactose is a disaccharide composed of both glucose and galactose and, while not wishing to be bound by any particular theory, is expected to follow a similar pattern to glucose. Similarly, mannose is a simple sugar known to be an isomer of glucose and, while not wishing to be bound by any particular theory, is thought to be metabolized via the same glycolytic pathway.

[0107] Glucose Patient selection All patients were recruited from St. Mary's Hospital between February 2019 and May 2019. Patients were recruited from three cohorts, namely esophageal cancer (n = 6), gastric cancer (n = 6) and age-matched healthy controls (n = 6). Written informed consent was obtained from all participants. Patients diagnosed with esophagogastric adenocarcinoma ranged from early-stage disease to metastatic palliative disease in the treatment pathway. Age-matched healthy controls included patients with benign upper gastrointestinal diseases (reflux, dysmotility) or healthy asymptomatic controls. Demographics and clinical information were collated.

[0108] Exhaled breath sampling​​​​​​​​​ A method for detecting four classes of volatile compounds, namely short-chain fatty acids, alcohols, aldehydes and phenol-alkanes was established by selected ion flow tube mass spectrometry (SIFT-MS Voi ceUltra 200; Syft Technologies, Anatune, UK ). All exhaled air sampling was performed in the morning, and patients maintained a clear liquid diet for at least 6 hours before exhaled air sampling. All patients exhaled directly into the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled air test was performed for each method, followed by consumption of 25 g of glucose dissolved in 100 ml of warm water, and then rinsing the mouth three times with water to decontaminate the oral cavity. Direct sampling of three exhaled air samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 1 5, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all four methods.

[0109] SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled air using chemical ionization. Precursor ions (H3O+, NO+ and O2+) are released into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled air is injected into the flow tube, reacted with the precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 - 30 °C.

[0110] Statistical analysis Data were obtained in parts per billion concentration units. SPSS statistical software (v25, Armo Using nk NY; IBM Corp), univariate analysis by Kruskal-Wallis was performed in 3 groups. The Mann-Whitney U test was performed to identify the differences between esophageal cancer and gastric cancer compared with the control. A P-value < 0.05 was considered statistically significant.

[0111] [Example 2] Tyrosine Patient Selection All patients were recruited from St. Mary's Hospital between February 2019 and May 2019. Patients were recruited from three cohorts: esophageal cancer (n = 6), gastric cancer (n = 6), and healthy age-matched controls (n = 6). Written informed consent was obtained from all participants. Patients diagnosed with esophagogastric adenocarcinoma ranged from early-stage disease to metastatic palliative disease in the treatment pathway. Healthy age-matched controls included patients with benign upper gastrointestinal diseases (reflux, dysmotility) or healthy asymptomatic controls. Demographics and clinical information were collated.

[0112] Exhaled Breath Sampling The detection method for four classes of volatile compounds, namely short-chain fatty acids, alcohols, aldehydes, and phenol-alkanes, was established by selected ion flow tube mass spectrometry (SIFT-MS Voi ceUltra 200; Syft Technologies, Anatune, UK ). Exhaled breath sampling was performed in the morning for all, and patients maintained a clear liquid diet for at least 6 hours before exhaled breath sampling. All patients exhaled directly into the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled breath test was performed for each method, followed by consumption of 2 g of tyrosine dissolved in 100 ml of warm water, followed by rinsing the mouth except ​​​​​​​​Rinsed the mouth three times with water for rinsing. Direct sampling of three exhaled breath samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes in all four ways (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes).

[0113] SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H3O+, NO+ and O2+) are emitted into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled breath is injected into the flow tube and reacted with the precursor ions to generate product ions, which are then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 - 30 °C.

[0114] Statistical analysis Data were obtained in parts per billion concentration units. Univariate analysis by Kruskal - Wallis was performed among three groups using SPSS statistical software (v25, Armonk NY; IBM Corp). The Mann - Whitney U test was performed to identify the differences between esophageal cancer and gastric cancer compared with the control. A P - value < 0.05 was considered statistically significant.

[0115] [Example 3] Phenylalanine Subject One healthy subject.

[0116] Dosage concentration: The recommended daily intake level recommended by the Food and Nutrition Board is 100 mg / kg per day for adults, and the maximum dosage is 3 g. [1] A single dose of 3 g was selected for this study.

[0117] ​​​​​​​ Exhaled breath sampling The detection methods for short-chain fatty acids, aldehydes, and phenol-alkanes were established by selected ion flow tube mass spectrometry (SIFT-MS VoiceUltra 200; Syft Technologies, Anatune, UK). All exhaled breath sampling was performed after a clear liquid diet for at least 6 hours in the morning. Exhaled breath was directed directly to the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled breath test was performed for each method, followed by consuming phenylalanine dissolved in 100 ml of warm water, followed by rinsing the mouth three times with water to decontaminate the oral cavity. Direct sampling of three exhaled breath samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all four methods.

[0118] SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H3O+, NO+, and O2+) are released into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled breath is injected into the flow tube, reacted with the precursor ions to produce product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 - 30 °C. Data were obtained in parts per billion concentration units.

[0119] [Example 4] Glutamic acid Subject One healthy subject.

[0120] Dosage concentration ​​​​​​​​The recommended daily intake level recommended by the Food Nutrition Committee is 30 mg / day for adults per kg. [1] A maximum single dose of 2.1 g was selected for an average 70 kg adult .

[0121] Exhaled breath sampling The detection methods for short-chain fatty acids, aldehydes and phenol-alkanes were established by selected ion flow tube mass spectrometry (SIFT-MS VoiceUltra 200; Syft Technologies, Anatune, UK). All exhaled breath sampling was performed after a clear liquid diet for at least 6 hours in the morning. Exhaled breath was directed directly to the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled breath test was performed for each method, followed by consuming glutamic acid dissolved in 100 ml of warm water, followed by rinsing the mouth 3 times with water to decontaminate the oral cavity. Direct sampling of 3 exhaled breath samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30 , 35, 40, 45, 50, 55, 60 minutes) for all 4 methods.

[0122] SIFT-MS SIFT-MS enables the real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H3O+, NO+ and O2+) are released into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled breath is injected into the flow tube, reacted with the precursor ions to produce product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 - 30 °C. Data were obtained in parts per billion concentration units.

[0123] ​​ [Example 5] Glycerol dosage Subjects Two healthy subjects volunteered to participate and gave written informed consent.

[0124] Dosage concentration Two dosages of the substrate were derived from (i) the recommended daily intake level by the Food and Nutrition Board and (ii ) the initial glucose method development test. The maximum recommended daily dosage is 276 mg / kg per day for adults, although there are no reports of harm from higher dosages. [1] For an average 7 0 kg individual, a maximum of 19 g of glycerol is recommended. Based on these findings, the present inventors selected dosages of 50 g, 25 g, and 10 g to compare the dosage response to glucose concentration. All findings were compared to a baseline of 0 g.

[0125] Exhaled air sampling The method for detecting short-chain fatty acids and aldehydes was established by selected ion flow tube mass spectrometry (SIFT-MS VoiceUltra 200; Syft Technologies, Anatun e, UK). All exhaled air sampling was performed in the morning, and the subjects maintained a clear liquid diet for at least 6 hours prior to exhaled air sampling. All subjects exhaled directly into the inlet of the SIFT-MS using a disposable mouthpiece. A baseline exhaled air test was performed for each method, followed by consumption of glycerol dissolved in 100 ml of warm water, and then rinsing the mouth 3 times with water to decontaminate the oral cavity. Direct sampling of 3 exhaled air samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all 4 methods.

[0126] ​​​​​ Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) SIFT-MS enables real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H3O+, NO+ and O2+) are released into a quadrupole mass filter and transported along a flow tube by an inert helium gas. Exhaled breath is introduced into the flow tube, reacted with the precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 to 30 °C. Data were obtained in parts per billion concentration units.

[0127] [Example 6] Glycerol Patient Selection All patients were recruited from St Mary's Hospital between February 2019 and December 2019. Patients were recruited from three cohorts: esophageal cancer (n = 6), gastric cancer (n = 6) and healthy age-matched controls (n = 6). Written informed consent was obtained from all participants. Patients diagnosed with esophagogastric adenocarcinoma ranged from early disease to metastatic palliative disease in the treatment pathway. Healthy age-matched controls included patients with benign upper gastrointestinal diseases (reflux, dysmotility) or healthy asymptomatic controls. Demographic and clinical information was collated.

[0128] Exhaled Breath Sampling The detection method for four classes of volatile compounds, namely short-chain fatty acids, alcohols, aldehydes and phenol-alkanes, was Selected Ion Flow Tube Mass Spectrometry (SIFT-MS Voi ceUltra 200; Syft Technologies, Anatune, UK ​​​​​​​​​​​​) was established. All exhaled air sampling was performed in the morning, and patients maintained a clear liquid diet for at least 6 hours before exhaled air sampling. All patients used disposable mouthpieces to exhale directly into the inlet of the SIFT-MS. A baseline exhaled air test was performed for each method, followed by consumption of 25 g of glycerol dissolved in 100 ml of warm water, and then rinsing the mouth 3 times with water to decontaminate the oral cavity. Direct sampling of 3 exhaled air samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all 4 methods. ring. All patients used disposable mouthpieces to exhale directly into the inlet of the SIFT-MS. A baseline exhaled air test was performed for each method, followed by consumption of 25 g of glycerol dissolved in 100 ml of warm water, and then rinsing the mouth 3 times with water to decontaminate the oral cavity. Direct sampling of 3 exhaled air samples over 60 seconds was performed continuously at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes) for all 4 methods. ring. ring.

[0129] SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled air using chemical ionization. Precursor ions (H O 3 O + NO + and O 2 + ) are released into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled air is injected into the flow tube, reacted with precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to an automated verification cycle once a day to operate within a temperature range of 10 - 30 °C. filter and transported along the flow tube by an inert helium gas. Exhaled air is injected into the flow tube, reacted with precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). filter and transported along the flow tube by an inert helium gas. Exhaled air is injected into the flow tube, reacted with precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to an automated verification cycle once a day to operate within a temperature range of 10 - 30 °C. filter and transported along the flow tube by an inert helium gas. Exhaled air is injected into the flow tube, reacted with precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to an automated verification cycle once a day to operate within a temperature range of 10 - 30 °C.

[0130] Statistical analysis Data were obtained in parts per billion concentration units. Univariate analysis by Kruskal-Wallis was performed among 3 groups using SPSS statistical software (v25, Armonk NY; IBM Corp). The Mann-Whitney U test was performed to identify the differences between esophageal cancer and gastric cancer compared with the control. A P value < 0.05 was considered statistically significant. groups using SPSS statistical software (v25, Armonk NY; IBM Corp). The Mann-Whitney U test was performed to identify the differences between esophageal cancer and gastric cancer compared with the control. A P value < 0.05 was considered statistically significant.

[0131] [Example 7] Compound amino acids (tyrosine, phenylalanine, glutamic acid) Patient selection All patients were recruited from St. Mary's Hospital between February 2019 and December 2019 Patients were recruited from three cohorts, namely esophageal cancer (n = 6), gastric cancer (n = 1) and healthy controls of the same age (n = 6). Written informed consent was obtained from all participants Patients diagnosed with esophagogastric adenocarcinoma ranged from early disease to metastatic palliative disease in the treatment pathway. Healthy controls of the same age included patients with benign upper gastrointestinal diseases (reflux, dysmotility) or healthy asymptomatic controls. Demographic and clinical information was collated.

[0132] Exhaled breath sampling The detection method for four classes of volatile compounds, namely short-chain fatty acids, alcohols, aldehydes and phenol-alkanes was established by selected ion flow tube mass spectrometry (SIFT-MS Voi ceUltra 200; Syft Technologies, Anatune, UK ). Exhaled breath sampling was performed in the morning for all patients, and patients maintained a clear liquid diet for at least 6 hours before exhaled breath sampling . All patients exhaled directly into the inlet of the SIFT-MS using a disposable mouthpiece . A baseline exhaled breath test was performed for each method, followed by consumption of 2 g of tyrosine, 3 g of phenylalanine dissolved in 100 ml of warm water and 2.1 g of glutamic acid, followed by rinsing the mouth three times with water to decontaminate the oral cavity . Direct sampling of three exhaled breath samples over 60 seconds was performed continuously for all four methods . Performed at 5-minute intervals for up to 60 minutes (0, 5, 10, 15, 20, 25, 30, 35, 40 , 45, 50, 55, 60 minutes).

[0133] SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H 3 O + , NO + and O 2 + ) are released into a quadrupole mass filter and transported along the flow tube by inert helium gas. Exhaled breath is injected into the flow tube, reacted with the precursor ions to generate product ions, and then separated according to the mass-to-charge ratio (m / z). SIFT-MS was subjected to an automated verification cycle once a day to operate within a temperature range of 10 - 30 °C.

[0134] Statistical analysis Data were obtained in parts per billion concentration units. SPSS statistical software (v25, Armonk NY; IBM Corp) was used to perform the Mann-Whitney U test between cancer and non-cancer. A P value < 0.05 was considered statistically significant.

[0135] [Example 8] Combination of glucose and citric acid Patient selection All patients were recruited from St. Mary's Hospital between February 2019 and December 2019. Twelve healthy controls were recruited to form two cohorts consuming glucose (n = 6) and a combination of glucose and citric acid (n = 6). Written informed consent was obtained from all participants. Healthy controls of the same age included patients with benign upper gastrointestinal diseases (reflux, dysmotility) or healthy asymptomatic controls. ​​​​​​​​​​

[0136] Exhaled gas sampling The detection method for four classes of volatile compounds, namely short-chain fatty acids, alcohols, aldehydes and phenol-alkanes was established by Selected Ion Flow Tube Mass Spectrometry (SIFT-MS Voi ceUltra 200; Syft Technologies, Anatune, UK ). Exhaled gas sampling was performed in the morning. Patients maintained a clear liquid diet for at least 6 hours before exhaled gas sampling. All patients exhaled directly into the inlet of the SIFT-MS using disposable mouthpieces. A baseline exhaled gas test was performed for each method, followed by consumption of 25 g of glucose and 1.4 g of citric acid dissolved in 100 ml of warm water, and then the mouth was rinsed three times with water to decontaminate the oral cavity. Direct sampling of three exhaled gas samples over 60 seconds was performed continuously at 5-minute intervals for up to 30 minutes for all four methods (0, 5, 10, 15, 20, 25, 30 minutes). SIFT-MS SIFT-MS enables real-time quantification and identification of VOCs in exhaled breath using chemical ionization. Precursor ions (H 3 O + , NO + and O 2 + ) are released into a quadrupole mass filter and transported along the flow tube by an inert helium gas. Exhaled breath is injected into the flow tube and reacted with the precursor ions to generate product ions, which are then separated according to the mass-to-charge ratio (m / z) . SIFT-MS was subjected to a daily automated verification cycle to operate within a temperature range of 10 - 30 °C.

[0137] Statistical analysis ​​Data was obtained in parts per billion concentration units. SPSS statistical software (v25, Armo nk NY; IBM Corp) was used to perform the Mann-Whitney U test between cancer and non-cancer. A P-value < 0.05 was considered statistically significant.

[0138] Results [Example 1] Glucose dosing test Analysis of volatile organic compounds

[0139]

Table 1

[0140] The increasing glucose concentration was positively correlated with the increasing concentration of volatile fatty acids detected in exhaled breath. Butyric acid and propanoic acid demonstrated a maximum response within 5 - 15 minutes of glucose consumption, after which the values decreased. Rapid glucose breakdown via the glycolytic pathway generates volatile end products detected in exhaled breath. Previous studies by the inventors have demonstrated that rinsing the mouth with water after glucose consumption eliminates potential VOC responses derived from the oral cavity. Butyric acid and pentanoic acid demonstrated a two-fold increase difference between 10 g and 50 g of glucose. Using these compounds, a recommended glucose dose for preclinical testing including patients with esophageal gastric cancer was derived. To obtain a balance between an appropriate dose response and a beverage tolerable to patients, the inventors selected a dose of 25 g dissolved in 100 ml of warm water. The next step in this study is to evaluate the VOC response in patients diagnosed with OG cancer compared to healthy age-matched controls and observe differences in cellular metabolic activity and VOC response.

[0141] ​​​​​​​​​​​[Table 2] JPEG2025081455000002.jpg126160

[0142]

Table 3

[0143] Glucose Analysis of Volatile Organic Compounds Short-chain Fatty Acids

[0144]

Table 4

[0145]

Table 5

[0146] Discussion Three chemical classes of VOCs in exhaled breath were demonstrated to be significantly different in patients diagnosed with esophageal-gastric (OG) cancer. A total of 13 compounds from the group, short-chain fatty acids (SCFAs) (n = 4), aldehydes (n = 6) and phenols (n = 3) demonstrated an increase in concentration after glucose consumption.

[0147] Volatile SCFAs, namely butanoic acid and propanoic acid, demonstrated the largest change in exhaled concentration. Optimal concentrations were reached within 10 minutes of consumption, suggesting rapid glycolysis. Glucose, a monosaccharide enters the glycolytic pathway that produces end-products of metabolism detected in exhaled breath. Pentanoic acid was detected at a higher concentration compared to baseline values at 30 minutes. These results suggest that exhaled VOCs can be increased by oral substrates by manipulating the specific metabolic pathways of known VOCs associated with OG cancer. Gastric cancer is detected in all ​​​A significant VOC demonstrated a stronger response than esophageal cancer. The gastric cancer group showed significant fold changes with SCFAs (acetic acid, butyric acid, pentanoic acid, and propanoic acid), and two of them had significant overlap with the esophageal cancer group (acetic acid and pentanoic acid).

[0148] Similarly, aldehydes such as pentanal and propanal followed a similar response pattern to SCFAs with an optimal concentration increase within 5 - 10 minutes. The remaining aldehydes consistently showed an increase in levels in the cancer groups, and 4 out of 9 had higher baseline values. Acetaldehyde, butanal, hexanal, and pentanal demonstrated a significant increase in fold change from the baseline in gastric cancer patients. Esophageal cancer patients showed this effect only with both butanal and propanal. On the other hand, for nonanal and octanal, both groups demonstrated a significant fold increase in the control group, which requires further exploration. These results are consistent with previous studies published by the inventors associating volatile butyric acid, butanal, and decanal with OG cancer. [1] The remaining aldehydes and phenol - alkanes (except dodecane) consistently showed an increase in concentration in the cancer groups over the duration of the study. Previous studies by the inventors' group have implied phenol as a potential breath biomarker in OG cancer. [2] Decane of the phenol family showed higher baseline concentrations in both cancer groups. The fold increase in the control group after glucose consumption requires further exploration. As a new finding, a similar response pattern was observed with p - cresol, but the fold increase was only seen in the gastric cancer group. This is in comparison to the retention of glucose in the stomach and the transient consumption of glucose by esophageal tumors These results are consistent with previous studies published by the inventors associating volatile butyric acid, butanal, and decanal with OG cancer. [1] The remaining aldehydes and phenol - alkanes (except dodecane) consistently showed an increase in concentration in the cancer groups over the duration of the study. Previous studies by the inventors' group have implied phenol as a potential breath biomarker in OG cancer. [2] These results are consistent with previous studies published by the inventors associating volatile butyric acid, butanal, and decanal with OG cancer. [1] The remaining aldehydes and phenol - alkanes (except dodecane) consistently showed an increase in concentration in the cancer groups over the duration of the study. Previous studies by the inventors' group have implied phenol as a potential breath biomarker in OG cancer. [2] As a new finding, a similar response pattern was observed with p - cresol, but the fold increase was only seen in the gastric cancer group. This is in comparison to the retention of glucose in the stomach and the transient This could reflect the passage of

[0149] Currently, the NICE guidelines state that there are "red flag" symptoms that may indicate OG cancer. The authors recommend that patients with these symptoms undergo upper gastrointestinal endoscopy within 2 weeks.[3] However, The insidious nature of the disease means that the majority present with non-specific symptoms, leading to delayed diagnosis and reduced overall survival. Noninvasive breath testing can help to stratify patients with nonspecific upper gastrointestinal symptoms. This will act as a triage tool to identify patients with OG cancer who have a history of cancer. Identification of these tumors has the potential to provide therapeutic treatment to patients and impact overall survival outcomes. The study evaluated patients with early and advanced stages of the disease.

[0150] In clinical practice, exhaled breath may be collected using: - A breath sampling device coupled with a thermal desorption tube to facilitate sample storage and transport .

[0151] - Direct sampling using mass spectrometry such as SIFT as demonstrated in this study .

[0152] - VOCs with large responses such as acetic acid, butanoic acid, pentanoic acid and propanoic acid Dedicated sensor for this purpose.

[0153] Main points Glucose consumption is a proxy for increased tumor microbiome or tumor cell activity. This activates the metabolic pathway, which can be detected by:

[0154] Significant fold increase in SCFAs (acetic, butanoic, pentanoic and propanoic acids). Esophagus This is observed more in the gastric cancer group than in the gastric cancer group.

[0155] · An increase in aldehydes, namely, acetaldehyde, butanal, hexanal and pentanal, was significant in the gastric cancer group. An increase in butanal and propanal was observed in esophageal cancer.

[0156] · New findings of an increase in the baseline concentrations of decane and p-cresol were observed in both cancer groups. The fold increase in p-cresol was shown only in gastric cancer.

[0157] Breath samples are collected at two intervals after glucose ingestion, namely, initially 5 - 10 minutes and later 30 minutes, to identify the optimal concentration of VOCs.

[0158] [Example 2] Tyrosine

[0159]

Table 6

[0160]

Table 7

[0161] Analysis of Volatile Organic Compounds Short-chain Fatty Acids

[0162]

Table 8

[0163] Aldehydes

[0164]

Table 9

[0165] Phenols

[0166] [Table 10]

[0167] [Table 11]

[0168] Consideration Two chemical classes of volatile compounds (phenols and aldehydes) were identified in tyrosine A total of eight compounds were detected in the esophageal cancer group at slightly increased concentrations 30 min after consumption. The underlying biological and mechanistic pathways are related to the Tyrosine, a fatty acid, is converted to a phenolic compound by an enzymatic reaction initiated by gastrointestinal bacteria. This suggests that it is metabolized to

[0169] Volatile phenolic compounds were detected at optimal concentrations 35–45 min after tyrosine consumption, A small increase from baseline was reported. Phenol and decane were similar between groups. The increase pattern was similar in the esophageal cancer group, whereas the concentrations of p-cresol and dodecane were significantly higher in the esophageal cancer group. Volatile aldehydes, namely butanal, decanal, , heptanal and hexanal demonstrated higher concentrations in the esophageal cancer group compared to the controls. (fold change 1.46 vs. 1.32). Overall baseline concentrations of all compounds were was significantly higher in the control group.

[0170] Decanal demonstrated the only significant fold increase in the esophageal cancer group, and reduced aldehyde dehydrogenase levels in OG cancer patients. The present study showed significantly higher baseline values ​​for aldehydes (butanal, decanal) and phenols. This is supported by previous studies by the inventors. [1, 2] The lack of support by the inventors' previous findings may be due to the fact that the results were obtained separately from the esophageal cancer group and the gastric cancer group and from a small number of patients, and further exploration is needed. However, the selected volatile compounds showed a response to tyrosine that was not significant but overall increased with fold change in cancer.

[0171] The short-chain fatty acid concentrations from the cancer cohort were not affected by tyrosine.

[0172] These results suggest the potential for exhaled VOCs to be increased by oral metabolic substrates acting via the shikimic acid pathway. In the next stage of the study, the inventors intend to use phenylalanine, a precursor of tyrosine in addition to tyrosine, as a combined beverage. Without wishing to be bound by any particular theory, the inventors aim to measure the exhaled VOC concentration between 30 and 45 minutes after ingestion and detect potential changes due to the addition of amino acids.

[0173] Highlights · Decanal of the aldehyde family demonstrated a significant fold increase after tyrosine consumption in the esophageal cancer group.

[0174] · Aldehyde and phenol compounds showed fold changes from a slightly increased baseline value that were not significant.

[0175] · Significantly higher baseline values of aldehydes and phenols in the control group need further exploration.

[0176] · Volatile phenol compounds were detected at optimal concentrations 35 - 45 minutes after tyrosine consumption.

[0177] [Example 3] Phenylalanine Results and Discussion Phenylalanine is an essential amino acid that is a known precursor of other amino acids such as tyrosine It is. Metabolism via the shikimic acid pathway is expected to produce volatile phenolic compounds Three compounds of the phenol family (dodecane, decane, and phenol) were shown to increase in concentration after phenylalanine consumption (Figs. 18-20). Dodecane and deca ne showed a maximum increase at 10-15 minutes after consumption (3.2- and 1.8-fold increases, respectively). Phenol showed a 2.7-fold increase at 60 minutes. Decanal and dodecane showed an increase in response to phenylalanine compared to tyrosine, which had no significant effect (Fig. 2 1). Phenol produced a similar final result, but decane showed a slightly increased value after phenylalanine intake .

[0178] [Example 4] Glutamic acid Results and Discussion Three compounds of the aldehyde and phenol families were shown to increase in VOC concentration after glutamic acid consumption (Figs. 22-25). Propanal showed the maximum increased concentration at 5 minutes, with a 3.5-fold change. Both dodecane and phenol showed a maximum 2-fold increase at 20 and 45 minutes, respectively. Glutamic acid is a non-essential amino acid that is metabolized via the shikimic acid pathway to produce volatile phenolic compounds. Glutamic acid is involved in the aminotransfer process during degradation. The resulting keto acid is used as an important intermediate in the citric acid cycle for further cell metabolism. This may explain the slight increase in butanoic acid observed within 5 minutes of glutamic acid consumption . ​​

[0179] While not wishing to be bound by any particular theory, the inventors have found that in combination with other amino acids tested, phenylalanine and tyrosine, an increased VOC response may occur, particularly with certain compounds already identified across the groups, namely dodecane, phenol. While not wishing to be bound by any particular theory, the inventors have found that in combination with other amino acids tested, phenylalanine and tyrosine, an increased VOC response may occur, particularly with certain compounds already identified across the groups, namely dodecane, phenol. While not wishing to be bound by any particular theory, the inventors have found that in combination with other amino acids tested, phenylalanine and tyrosine, an increased VOC response may occur, particularly with certain compounds already identified across the groups, namely dodecane, phenol. While not wishing to be bound by any particular theory, the inventors have found that in combination with other amino acids tested, phenylalanine and tyrosine, an increased VOC response may occur, particularly with certain compounds already identified across the groups, namely dodecane, phenol.

[0180] [Example 5] Glycerol dosage Results Subject Two subjects with an average age of 32 years were recruited, namely 1 female and 1 male. No significant co-existing diseases were observed. Two subjects with an average age of 32 years were recruited, namely 1 female and 1 male. No significant co-existing diseases were observed.

[0181] Analysis of Volatile Organic Compounds

[0182] [Table 12]

[0183] Discussion The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors The increasing concentration of glycerol leads to an increase in the production of volatile fatty acids detected in exhaled breath. The concentration of volatile fatty acids from 25 g of glycerol is equivalent to the baseline value. The concentration of butyric acid rises 30 minutes after glycerol intake, and the maximum concentration is detected at 45 - 55 minutes. Glycerol is a polyol compound found in lipids and is metabolized either (i) through the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of maximum fatty acids 5 - 10 minutes after glucose consumption, and thus the response after glycerol intake is expected to be delayed as further enzymatic reactions may be required before entering the circuit. The inventors Intended to use a 50 g dose to elicit the fatty acid VOC response in the breath of patients diagnosed with OG cancer, compared to healthy age-matched controls.

[0184] [Example 6] Glycerol Results Patients Eighteen patients were recruited (n = 6 in each group; esophageal cancer, gastric cancer, healthy controls). All cancers included were histologically confirmed as adenocarcinoma.

[0185] [Table 13]

[0186] Analysis of Volatile Organic Compounds Short-chain fatty acids

[0187] [Table 14]

[0188] Aldehydes

[0189] [Table 15]

[0190] Phenols

[0191] [Table 16]

[0192] [Table 17]

[0193] Discussion Three classes of chemical substances of VOCs in exhaled breath were demonstrated to increase significantly in patients diagnosed with esophagogastric (OG) cancer. Glycerol is a polyol compound found in lipids and is (i) metabolized via the glycolytic pathway by directly entering the glycolytic pathway or (ii) converted to glucose by gluconeogenesis. The glucose test demonstrated the detection of the maximum fatty acids 5 - 10 minutes after glucose consumption. Therefore, the increase in response after glycerol intake is expected to be further delayed because an enzymatic reaction may be required before entry into the circuit and may be further delayed.

[0194] Consistent with the hypothesis, the inventors observed an increase in the levels of short - chain fatty acids (SCFAs) and aldehydes between 45 - 60 minutes after glycerol consumption, as shown in Figures 30A - 30E. SCFAs, namely acetic acid, butyric acid, and propanoic acid, showed a greater increase in the esophageal cancer group (1.5, 2.02, 1.75 - fold increases respectively) compared to the gastric cancer group (1.09, 1.43, 1.46 - fold increases respectively). A gradual increase was observed after 45 minutes and the optimal concentration was reached at 60 minutes.

[0195] Similarly, the selected aldehydes were found to increase significantly in the esophageal cancer group (Figures 31A - 31I). Hexanal and propanal showed the largest increase with a 1.7 - fold increase between 40 - 55 minutes. Octanal increased with a 1.58 - fold change and pentanal increased with a 1.27 - fold change. The gastric cancer group showed a change only due to a 1.46 - fold increase in pentanal at 55 minutes. The remaining aldehydes were not affected.

[0196] Multiple volatile phenols tested showed differences in exhaled breath concentrations among the three patient groups after glycerol consumption. ​​​​​​​​No significant change was demonstrated (Figs. 32A - 32E).

[0197] Glycerol consumption uniquely increased the target VOCs in the esophageal cancer group. This may be due to an increase in the viscosity of the fluid covering the esophagus, allowing for transient or greater passage. An increase in the contact time between the substrate and the tumor can explain the occurrence of the increased VOC levels.

[0198] Highlights Glycerol consumption activates the glycolytic pathway associated with an increase in the activity of the tumor microbiome or tumor cells. This is detected by the following.

[0199] · A significant fold increase in SCFAs (acetic acid, butyric acid, and propanoic acid). This is more pronounced in the esophageal cancer group than in the gastric cancer group.

[0200] · A significant increase in aldehydes in the esophageal cancer group, namely hexanal, octanal, pentanal and propanal. An increase in pentanal was observed in gastric cancer.

[0201] [Example 7] Composite amino acids (tyrosine, phenylalanine, glutamic acid) Results Patients Thirteen patients were recruited (esophageal cancer n = 6, gastric cancer n = 1, healthy controls n = 6). All cancers included were histologically confirmed as adenocarcinoma.

[0202] [Table 18]

[0203] Analysis of volatile organic compounds Short-chain fatty acids

[0204] ​​

Table 19

[0205] Aldehyde

[0206]

Table 20

[0207] Phenol - alkane

[0208]

Table 21

[0209] Discussion Two classes of chemicals, aldehydes and phenol - alkanes, demonstrated an increase in volatile organic compound levels after consumption of three composite amino acids, as shown in Figures 33 and 3 4A - 34E. In contrast, when tyrosine alone was administered, only decanal slightly increased in the esophagus group . The aldehyde decanal demonstrated a more significant increase in detection levels with this amino acid combination beverage (Figure 33). An increase of 1.41 - fold was observed in the cancer group compared to a 1.05 - fold increase in the control group (baseline = 0.69 ppbv, 30 minutes = 0

[0210] .83 ppbv). The maximum concentration occurred 30 minutes after consumption of the nutrient beverage . Phenol - alkanes are a major target of this nutrient group. The pathway explaining the metabolism of tyrosine by tyrosine phenol lyase to produce phenol is detailed .

[0211] . More recently, Saito et al. described an enzyme tyrosine lyase for producing p - cresol . . It describes the pathway involving metabolism by Ze. This metabolic pathway has been demonstrated in bacteria [4], rather than in human cells. p-Cresol significantly increased from a baseline level of 0.93 ppbv to 1.25 ppbv 40 minutes after consumption of the amino acid beverage, leading to a 1.37-fold increase. No change was observed in the control group. Phenol showed an overall increase both in the cancer group (a 1.79-fold increase) and the non-cancer group (a 1.83-fold increase), with no significant difference between the two. Decanal also increased at 30 minutes, resulting in a 1.44-fold increase in the cancer group. There was no significant change in the remaining aldehydes and short-chain fatty acids. Further investigation is needed to explain the increase in decanal. .93 ppbv baseline level to 1.25 ppbv and led to a 1.37 -fold increase. No change was observed in the control group. Phenol showed an overall increase both in the cancer group (1 .79-fold increase) and the non-cancer group (1.83-fold increase), with no significant difference between the two. Decanal also increased at 30 minutes and resulted in a 1.44 -fold increase in the cancer group.

[0212] There was no significant change in the remaining aldehydes and short-chain fatty acids. Further investigation is needed to explain the increase in decanal.

[0213] Highlights: Consumption of the complex amino acids potentially activates metabolic pathways related to bacteria. This is detected by the following.

[0214] · A significant fold increase in decanal (aldehyde).

[0215] · An overall increase in phenol (enzyme tyrosine phenol lyase ) without a difference between the cancer group and the non-cancer group.

[0216] · A new finding of a significant increase in p-cresol potentially caused by enzyme metabolism using tyrosine lyase.

[0217] [Example 8] Combination of glucose and citric acid

[0218] [Table 22]

[0219] Analysis of Volatile Organic Compounds Short-chain Fatty Acids

[0220]

Table 23

[0221] Aldehydes

[0222]

Table 24

[0223] Discussion Two classes of chemicals, short-chain fatty acids (SCFAs) and aldehydes, were shown to increase volatile organic compound levels after consumption of a combination of glucose and citrate, as can be seen in Figures 35A - 3 5E and 36. The results of Example 1 demonstrated a significant increase in these groups within 5 - 10 minutes of glucose consumption alone. According to the hypothesis, glucose is metabolized through the glycolytic pathway that occurs in human cells and bacterial cells. Since the glycolytic pathway feeds into the citric acid cycle, the aim was to evaluate the further increase in VOCs due to the addition of citrate. Butyric acid demonstrated the largest increase, from 2.72 - fold with glucose alone to 5.36 - fold with the addition of citrate (Figure 35B). The maximum concentration was achieved within 5 - 10 minutes of beverage consumption. Propionic acid also demonstrated a 1.96 - fold increase with the addition of citrate (Figure 35 E). No significant changes were observed in the remaining SCFA groups. Propanal showed a slight change, but a 1.29 - fold increase in the citrate group within 10 - 15 minutes.

[0224] Butyric acid demonstrated the largest increase, from 2.72 - fold with glucose alone to 5.36 - fold with the addition of citrate (Figure 35B). The maximum concentration was achieved within 5 - 10 minutes of beverage consumption. Propionic acid also demonstrated a 1.96 - fold increase with the addition of citrate (Figure 35 E). No significant changes were observed in the remaining SCFA groups. Propanal showed a slight change, but a 1.29 - fold increase in the citrate group within 10 - 15 minutes. No significant changes were observed in the remaining SCFA groups.

[0225] Propanal showed a slight change, but a 1.29 - fold increase in the citrate group within 10 - 15 minutes. ​It seemed to be the only aldehyde showing an increase (Figure 36). No significant changes were observed in the remaining aldehyde groups. No significant changes were observed.

[0226] Using citrate as a differentiating factor, clear changes in VOCs were shown in two control groups. The present inventors hypothesize that these nutrients may be supplied to the specific glycolysis and citrate metabolic pathways. The present inventors hypothesize that these nutrients may be supplied to the specific glycolysis and citrate metabolic pathways. Hypothesize.

[0227] Highlights The consumption of the combination of glucose and citrate activates known metabolic pathways related to cell metabolism. This is detected by the following: This is detected by the following. · A significant fold increase in volatile short-chain fatty acids (butyric acid and propanoic acid) within 5 - 10 minutes of consuming the nutrient beverage. ). · A significant fold increase in propanal within 10 - 15 minutes. The next step involves recruiting patients with oesophagogastric cancer and evaluating the changes in exhaled VOC in response to additional nutrient substrates. including evaluating changes in exhaled VOC in response to additional nutrient substrates.

[0228] References 1. Markar, S.R., et al., Assessment of a Noninvasive Exhaled Breath Test for the Diagnosis of Oesophagogastric Cancer. JAMA Oncol, 2018. 4(7): p. 970 - 976. 2. Kumar K, H.J., Abbassi - Ghadi N, Mackenzie HA, Veselkov KA, Hoare JM, Lovat L B, Spanel P, Smith D and Hanna GB, Mass Spectrometric Analysis of Exhaled Breath for the Identification of Volatile Organic Compound Biomarkers in Esophageal an d Gastric Adenocarcinoma. Annals of Surgery, 2015. 262(6): p. 981-990. 3. Excellence, N.I.o.C., Gastrointestinal tract (upper) cancers - recognition an d referral. 2016. 4. Saito Y, Sato T, Nomoto K, Tsuji H. Identification of phenol- and p-cresol- p roducing intestinal bacteria by using media supplemented with tyrosine and its m etabolites. FEMS Microbiol Ecol. 2018. 94(9)

Claims

1. To diagnose a subject suffering from or predisposed to cancer, or to provide a prognosis of said subject's condition. A method for producing a medicament for use in a medical device, comprising: (i) detecting in a body sample from a test subject a small amount of IgG present in a composition previously administered to said subject; At least one sugar and / or at least one amino acid or a precursor thereof and / or or a detector for detecting the concentration of a trace compound produced by metabolism of at least one polyol. wherein the sugar is present in the composition at a concentration greater than 20,000 mg / 100 ml. and the amino acid or precursor thereof is present in the composition at a concentration of at least 500 mg / ml. and the polyol is present in the composition at a concentration of greater than 25,000 mg / 100 ml. detecting the presence of the compound in an object; and (ii) comparing this concentration to a baseline concentration of said trace compound in individuals not suffering from cancer. Step an increase or decrease in the concentration of the trace compound compared to the reference indicates that the subject suggesting that the subject suffers from or has a predisposition to cancer, or denying the subject's condition A method for providing a predictive prognosis.

2. The detection step (i) comprises detecting the trace compound by detecting at least one sugar and / or amino acid. or a precursor thereof and / or from the administration of said composition comprising at least one polyol. Check within 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, or 5 minutes. The method of claim 1 , further comprising:

3. The detection step (i) comprises detecting the trace compound by detecting at least one sugar and / or amino acid. or a precursor thereof and / or from the administration of said composition comprising at least one polyol. Between 30 and 60 minutes, or between 30 and 55 minutes, or between 30 and 50 minutes, or between 30 and Between 45 minutes, or between 30 and 40 minutes, or between 35 and 60 minutes, or between 35 and 55 minutes Detected between 35 and 50 minutes, between 35 and 45 minutes, or between 35 and 40 minutes The method of claim 1 or 2, comprising:

4. An increase in the concentration of the trace compound compared to the standard is an indication that the subject may be suffering from cancer. or suggest a predisposition thereto, or provide a negative prognosis of the subject's condition. The method according to any one of claims 1 to 3,

5. The increase in the concentration of the trace compound is at least 1% of the increase in the concentration of the trace compound compared to the reference. ０％、２０％、３０％、４０％、５０％、１００％、２００％、３００％、４００％、５ 00%, 600%, 700%, 800%, 900% or 1000% increase in claims Item 5. The method according to item 4.

6. The sugar is present in the composition previously administered to the subject in an amount of at least 20,500 mg / ml. The method of any one of claims 1 to 5, which is present in a concentration of 100 ml.

7. The sugar is glucose, sorbitol, mannose or lactose.

7. The method according to any one of claims 6 to 6.

8. The sugar is glucose, and at least two of the sugars in the composition have been previously administered to the subject. 5,000 mg / 100 ml, and said trace compounds include glucose. The method of any one of claims 1 to 7, wherein detection occurs up to 10 minutes after administration of the composition.

9. The composition administered to the subject comprises citric acid in combination with the sugar, The acid is present in the composition in a concentration of at least 1,000 mg / 100 ml, and optionally The method according to any one of claims 1 to 8, wherein the sugar is glucose.

10. The amino acid is tyrosine, glutamic acid, glutamate, phenylalanine, triglyceride, is selected from the group consisting of heptaphan, proline and histidine, and optionally, said composition 10. The method according to claim 1, comprising administering to said subject a compound comprising tyrosine, phenylalanine and glutamic acid. The method described.

11. the amino acid is tyrosine, and the composition previously administered to the subject contains at least and optionally, said trace compounds include tyrosine. The method of claim 10, wherein the detection is performed between 35 and 45 minutes after administration of the composition.

12. The amino acid precursor is phenylalanine, and optionally at least 3000 mg / 10 12. The method of any one of claims 1 to 11, wherein the composition is present in a concentration of 0 ml.

13. The method according to any one of claims 1 to 12, wherein the polyol is glycerol.

14. The polyol is present in the composition at a concentration greater than 30,000 mg / 100 ml. The method according to any one of claims 1 to 13,

15. The cancer is esophagogastric junction cancer, gastric cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), Or esophageal adenocarcinoma (EAC).

16. Any of claims 1 to 15, wherein the cancer is gastric cancer, esophageal cancer or metastatic cancer.

2. The method according to claim 1.

17. The trace compounds are short chain fatty acids, aldehydes, alcohols, or any combination thereof. The method according to any one of claims 1 to 16.

18. The trace compounds are selected from the group consisting of C1-C3 aldehydes, C1-C3 alcohols, and C2-C10 alkanes substituted with an ═O group and the second carbon atom substituted with an —OH group; C1 ~C20 alkanes, C4-C10 alcohols, C1-C6 carboxylic acids, C4-C20 alkane aldehyde, a phenol optionally substituted with a C1-C6 alkyl group, a C2 aldehyde, 3 aldehyde, C8 aldehyde, C9 aldehyde, C10 aldehyde, C11 aldehyde , an analog or derivative of the aforementioned species, or any combination thereof. The method described.

19. The trace compounds are acetic acid, butanoic acid, hexanoic acid, pentanoic acid, propanoic acid, acetone, Aldehyde, decanal, heptanal, hexanal, nonanal, octanal, pentanal Tanal, butanal, propanal, 1-hydroxy-4-ethylbenzene, decane, dodecane, p-cresol and phenol or any combination thereof 19. The method of claim 17 or 18, wherein

20. The substrate is a sugar, preferably glucose, and the trace compounds are acetic acid, butanoic acid, Pentanoic acid, propanoic acid, hexanoic acid, acetaldehyde, propanal, butanal, Hexanal, pentanal, decanal, 1-hydroxyethylbenzene and / or The method according to any one of claims 17 to 19, wherein is p-cresol.

21. The substrate is an amino acid or a precursor thereof, and the trace compound is butanal, decanal, aldehyde, heptanal, hexanal, phenol, decane, p-cresol, 1-hydroxy 20. The method according to claim 17, wherein the diethylbenzene and / or dodecane are Method of posting.

22. the amino acid or precursor thereof is tyrosine, and the trace compound is decanal and The method according to claim 21 , wherein the aryl group is dodecane or dodecane.

23. The substrate is a polyol, preferably glycerol, and the trace compound is butanoic acid, Acetic acid, hexanoic acid, pentanoic acid, propanoic acid, butanal, hexanal, pentanal and / or propanal.

24. 1. A method for detecting a trace compound in a test subject, comprising: (i) subjecting the target to at least one of the methods according to any one of claims 1 to 23, which results in a trace compound; Providing a composition comprising a substrate; and (ii) detecting the concentration of said trace compound in a body sample from said subject; A method comprising:

25. 2. The method of claim 1, wherein the trace compound is as defined in any one of claims 17 to 23.

5. The method according to claim 4.

26. A small amount of the present compound is suitable for metabolism into trace compounds for use in diagnostic or prognostic methods. At least one sugar and / or at least one amino acid or its precursor and / or or at least one polyol, wherein the sugar is 20,000 mg / l. 00ml, and the amino acid is present in the composition at a concentration of at least 500 mg / ml. ml, and the polyol is present in the composition at a concentration of 25,000 mg / 100 ml or more. The composition of claim 1, wherein the compound is present in said composition at a concentration greater than or equal to 100% by mass.

27. A method for detecting cancer by isolating a compound present in the present invention, the compound being suitable for metabolism into a trace compound for use in a method for the diagnosis or prognosis of cancer. at least one sugar and / or at least one amino acid or a precursor thereof; and / or at least one polyol, wherein the sugar is 20,000m g / 100 ml of the composition, and the amino acid is present in the composition at a concentration of at least 500 and the polyol is present in the composition at a concentration of 25,000 mg / 100 ml. and optionally, the cancer is gastroesophageal junction cancer, stomach cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), or esophageal adenocarcinoma (EAC). Composition.

28. A method according to any one of claims 1 to 14 for use in a method according to any one of claims 1 to 23. A composition comprising at least one substrate according to any one of claims 1 to 4.

29. For diagnosing a subject suffering from or predisposition to cancer, or for providing a prognosis of said subject's condition. A kit for providing (a) a composition comprising at least one substrate as defined in any one of claims 1 to 14, (b) a means for determining the concentration of the trace compound in a sample from a test subject; and (c) a standard of concentrations of said trace compounds in samples from individuals not afflicted with cancer; a concentration of said trace compound in a body sample from said test subject compared to said standard; and identifying an increase or decrease, thereby determining whether the subject has or is predisposed to having cancer. or to provide a negative prognosis of the subject's condition. ,kit.

30. The trace compounds are as defined in any one of claims 17 to 23.

30. The kit of claim 29.

31. Subjects with cancer treated with therapeutic drugs or special diets, or with chemotherapy or chemoradiotherapy 1. A method for determining the efficacy of treatment of (i) the subject comprises at least one substrate according to any one of claims 1 to 14; Providing a composition; and (ii) a precursor produced by metabolism of said at least one substrate in a body sample from a test subject; The concentration of the trace compound is analyzed and the concentration is compared to the concentration of the trace compound in individuals not suffering from cancer. comparing the concentration of the compound to a standard; a concentration of said trace compound in said body sample from said test subject compared to said standard; The increase or decrease in the degree of the disease is due to the therapeutic agent or the special diet, or due to chemotherapy or chemical radiation. A method that indicates that a radiotherapy treatment regime is effective or ineffective.

32. The trace compound is as defined in any one of claims 17 to 23.

31. The method according to claim 31.

33. The cancer is esophagogastric junction cancer, gastric cancer, esophageal cancer, esophageal squamous cell carcinoma (ESCC), or esophageal adenocarcinoma (EAC).

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